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Manning throws 3 TDs as Broncos top Bucs 31-23
December 2, 2012
DENVER (AP) — Ah, that growing list of Broncos playmakers: Peyton Manning. Von Miller. Demaryius Thomas.
Mitch Unrein?
Well, yes.
The second-year defensive lineman out of Wyoming played as big a role as Manning, Miller or anyone else Sunday in helping the Denver Broncos defeat the Tampa Bay Buccaneers 31-23 and wrap up their second straight AFC West title.
Unrein caught a 1-yard touchdown pass from Manning to open the scoring, then got the hit on Tampa Bay quarterback Josh Freeman that forced a bad throw. It ended up in Miller's hands and, moments later, the end zone for the score that turned the game into a runaway.
"People are going to have to game-plan for that guy," cornerback Champ Bailey said.
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Thomas caught two other touchdown passes from Manning, who now has 29 this year and passed Jake Plummer and John Elway for the most scoring throws in a single season.
The Broncos (9-3) won their seventh straight and wrapped up a date in January in the playoffs, though there were no champagne bottles being uncorked in the locker room.
"It's one step," Bailey said. "It's not like we've done everything we want to do."
The Bucs (6-6), surprising contenders under first-year coach Greg Schiano, fell a game behind Seattle for the second NFC wild-card spot and their loss clinched the NFC South for Atlanta. They were in this game until Unrein made his second big play and set the stage for Miller's first NFL touchdown, which capped a 21-0 third quarter for Denver.
"The reality is, we get hit and throw the pick-6," Schiano said. "That changes the complexion of the game. Now we're not able to run the football. I mean, we didn't run it gang-busters in the first half. But we ran it. And … if you're able to be in a game where you can run it, oftentimes you'll be able to run it in the fourth quarter."
The Broncos held Tampa Bay rookie Doug Martin to 56 yards on 18 carries.
Manning went 27 for 38 for 242 yards, as the Broncos scored 30-plus points for the sixth time over their seven-game win streak — a night-and-day difference from last year, when they won the division at 8-8 and played close-to-the-vest, low-scoring games with Tim Tebow running the offense.
Yet despite Denver's big numbers Sunday, which included eight catches for 99 yards for Thomas and nine receptions for 89 yards from tight end Jacob Tamme, this was, yet again, a far-from-perfect effort from Manning and Co.
The quarterback was frustrated in this first half, as a series of penalties, off-target throws and missed protections kept him biding his time on the bench. The only real highlight came at the end of his first drive — a 1-yard touchdown pass to Unrein, who became the first defensive lineman to catch a touchdown pass in Broncos history, then celebrated with a big-time spike.
"After I spiked it, I was, like, 'Uh, oh, where'd it go?" Unrein said. "Luckily, I got it back."
Unrein's hit on Freeman in the third quarter also resulted in a touchdown — this time, from Miller, who went 26 yards to make it 28-10. It was Miller's first career score and he celebrated by doing his dance thing, not in the backfield, but this time in the other team's end zone.
"When I got my hands on it, I felt like I only had some O-linemen in front of me," Miller said. "I felt like my odds were pretty good."
Before the Miller TD, this was a game.
Late in the first quarter, Freeman found Mike Williams in single coverage down the middle for a 40-yard gain. Martin followed with a 7-yard run, then another for 5, then Freeman found Manning's old buddy in Indianapolis, Dallas Clark, for an 11-yard touchdown and a 10-7 lead.
But when the teams came out after halftime, Denver was a different team.
"The hype these guys get, it's well deserved," Freeman said. "They get after it. The front four, they do a good job of timing their blitzes. The Denver defense played a great game today."
Manning wasn't bad, either. With his ninth completion, he surpassed Dan Marino for second on the all-time completions list. He now has 4,986.
On the second drive of the third quarter, Manning completed three passes on a quick, 52-yard touchdown drive for a 14-10 lead, capping it with an 8-yard pass to Thomas, who had a sliver of space in the back of the end zone.
After a three-and-out, Manning did it again, starting a 57-yard drive with a 14-yard pass to Tamme and capping it five plays later with a 10-yard score to Thomas, who beat double coverage.
"What I liked about the second touchdown was that Demaryius was seeing the same thing I was seeing," Manning said. "We had another route on. We changed it at the line. We were thinking along the same lines. I like to see the communication, the growth."
After the early touchdown drive, Freeman never got comfortable again.
He finished 18 for 39 for 242 yards. He led the Bucs on a 47-yard touchdown drive that made the score 31-23 with 2:33 left. Denver recovered the onside kick and lined up in victory formation twice. Schiano, as is his wont, had his players challenge those plays, which led to some scrapping, but no coach-to-coach conflict the way Giants coach Tom Coughlin confronted Schiano under similar circumstances earlier this year.
"We were prepared for it," Broncos coach John Fox said.
Notes: Broncos K Matt Prater missed from 47 yards, making him 2 for 5 from 40 to 49 yards this season and 15 for 17 from everywhere else, including 3 for 4 from 50-plus. … Martin was held under 60 yards for the second straight week. … Matthew Willis, getting playing time that would've gone to injured Brandon Stokley, had three catches for 22 yards.


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Dr. Elise A. Feingold is a Program Director in Genome Analysis at the National Human Genome Research Institute (NHGRI). She received a B.A. in biology from the University of Pennsylvania and a Ph.D. in Human Genetics from Yale University. She completed post-doctoral research at the National Heart, Lung, and Blood Institute.
Following a one-year training experience in National Institutes of Health (NIH) extramural programs as a grants associate, Dr. Feingold joined the extramural division of what is now the National Human Genome Research Institute in 1992. Since that time, her area of focus has been on gene discovery, gene expression and functional genomics.
From its inception until 2004, Dr. Feingold managed, in collaboration with the National Cancer Institute, the Mammalian Gene Collection project, which created a comprehensive collection of sequence verified full-length human and mouse cDNA clones.
Dr. Feingold currently leads the Encyclopedia of DNA Elements (ENCODE) and the related project in model organisms (modENCODE) to identify all of the functional elements in the human, and the D. melanogaster and C. elegans genomes, respectively. In addition, Dr. Feingold manages a portfolio of research grants in functional genomics including several Centers of Excellence in Genomic Sciences (CEGS). Dr. Feingold serves on the NIH Common Fund Epigenomics and High Risk - High Reward working groups.
Last Reviewed: September 17, 2018

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In 1842, the shoe and leather industry was an important source of jobs and income for may people in Massachusetts. This convention was called because of the concern about the lack of tariffs that would help prohibit cheap imports. In 1832, a tariff act was passed which assessed specific rates of taxes to imports. This very unpopular tariff was modified by the Compromise Tariff of 1833, which lowered the tariff rates over a period of ten years to the point where most goods would be taxed at 20%. As this rate approached, members of the Whig Party called for protection from competition from European manufacturers. The speaker quoted here cites prices of goods, as well as claiming that the workers in Europe do not make a living wage, but are subsidized. The Tariff of 1842 was finally passed, and restored protection from cheap European goods by raising the rates to around 40%.
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People of African descent have been a part of Virginia's-and America's-story since European colonization of the continent began. Yet the contributions of African Americans have often been ignored, obscured, or under appreciated by those who recorded history. In observance of African American History Month, the Library of Virginia is pleased to honor eight distinguished Virginians as African American Trailblazers for their contributions to the state and nation.
The men and women featured as Trailblazers offer powerful examples of individuals who refused to be defined by their circumstances. Their biographies are a testament to the determination and perseverance displayed by extraordinary people during challenging times. Through education and advocacy, these individuals demonstrate how African Americans have actively campaigned for better lives for themselves and their people. It is these many contributions that the African American Trailblazers program seeks to share.
Below is the current year's trailblazers. A listing of past and present trailblazers is available alphabetically or listed by year.
Check The Richmond Times-Dispatch each Tuesday and Thursday during February for African American Trailblazer profiles.
2012 Program:
2012 African American Trailblazers: Program and Reception
Thursday, February 23, 2012
Time: 6:00 PM-8:00 PM
Place: Lecture Hall & Lobby
Free but reservations required. Seating is limited. Call 804-692-3535 by February 17 to RSVP. Eight honorees-from the past and present-who have had a significant impact on the history of Virginia will be honored at this celebration. A reception follows the program. Sponsored by Dominion.
2012 Trailblazers:
Michael L. Blakey (February 23, 1953– ), Williamsburg
Physical anthropologist Michael L. Blakey helped analyze and interpret the important evidence uncovered at the African Burial Ground in Manhattan
Noah Davis (March, 1804–April 7, 1867), Fredericksburg
author of an emancipation narrative
Born into slavery, Noah Davis raised more than $4,000 to free himself and his family members during the 1840s and 1850s.
James Farmer (1920–1999), Spotsylvania County
Civil rights leader
As director of the Congress of Racial Equality and as initiator of the Freedom Rides, James Farmer played a critical role in the national civil rights movement.
John Jasper (1812–1901), Richmond
Baptist minister
An extraordinary orator who preached throughout the eastern United States, John Jasper in 1867 established Sixth Mount Zion Baptist Church, in Richmond.
Willie Lanier (August 21, 1945– ), Richmond
Athlete and entrepreneur
Willie Lanier broke through racial barriers in professional football by becoming the first African American to play middle linebacker, the position that directs the defense on the field.
Oscar Micheaux (1884–1951), Roanoke
Often recognized as the country's first African American filmmaker, Oscar Micheaux established a film office in Roanoke, where he produced at least six silent movies.
Yvonne B. Miller (July 4, 1934–July 3, 2012), Norfolk
Political leader
The first African American woman elected to the House of Delegates and to the Senate of Virginia, Yvonne B. Miller worked tirelessly as a politician, educator, and advocate for underrepresented Virginians.
Irene Amos Morgan (1917–2007), Gloucester County
Principal in a civil rights case
Irene Morgan's challenge to the Virginia law requiring segregated seating on interstate buses resulted in a landmark ruling by the Supreme Court of the United States.
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The Fire's dip back below the red line on Saturday night was one of two moves in the Eastern Conference playoff race. With just over a month to play, certain teams are circling in on a playoff berth while others are battling for position.
The latest edition of Playoff Math awaits you below:
1) New York Red Bulls
Current Points: 51 (15-9-6)
Games Remaining: 4 (2 home / 2 away)
Last 5 Games: WWWWL (12pts)
2013 Home/Away Record: 10-2-3 / 5-7-3
Maximum Possible Pts: 63
Average Opponents Pts: 43.0
Remaining Schedule: 9/29 @ SEA, 10/5 vs. NE, 10/20 @ HOU, 10/27 vs. CHI
Rundown: New York watched Montreal lose and Sporting KC win on Saturday, knowing a home game against FC Dallas awaited the following day. It wasn't a great match but New York utilized a 76th minute own goal from Erick to earn another home result and somewhat quietly move into first place in the Supporters Shield race in the process.
The Red Bulls continue to benefit from the two teams chasing them having Champions League matches but a top of the conference clash awaits them Sunday night when they visit Seattle in a match that could go a long way to decide the first silverware of the season.
A New York win or tie Sunday combined with Fire and Philadelphia Union losses would see the Red Bulls become the first team to qualify for the postseason.
2) Sporting KC
Current Points: 48 (14-9-6)
Games Remaining: 5 (2 home / 3 away)
Last 5: WWWLL (9pts)
2013 Home/Away Record: 8-4-3 / 6-5-3
Maximum Possible Pts: 63
Average Opponents Pts: 34.8
Remaining Schedule: 9/27 vs. PHI, 10/5 @ CLB, 10/9 at HOU, 10/18 vs. D.C., 10/23 vs. OLI**, 10/26 @ PHI
Rundown: Despite a scare, Sporting earned a point against Real Esteli in Champions League play at midweek and will just need another draw vs. Olimpia to guarantee final passage. In MLS play, they fought hard for a 2-1 win at Toronto took benefit of Montreal's defeat to Vancouver to move into sole possession of second place in the East.
Sporting holds a game in hand over Red Bull but faces three matches against teams fighting for their playoff fate over the next two and a half weeks. Though their remaining schedule is easiest in the East, it will be a battle to stay in second and potentially push for the top spot.
3) Montreal Impact
Current Points: 45 (13-9-6)
Games Remaining: 6 (2 home/ 4 away)
Last 5 Games: LLWDW (7pts)
2013 Home/Away Record: 9-3-3 / 4-6-3
Maximum Possible Pts: 63
Average Opponents Pts: 38.2
Remaining Schedule: 9/24 vs. HER** 9/28 @ CHI, 10/4 @ HOU, 10/12 vs. NE, 10/16 @ LA, 10/19 vs. PHI, 10/26 @ TFC
Rundown: So what's going on with Montreal? Their 3-0 loss to Vancouver marked their third straight across all competitions and second straight at home in MLS. Like their performances this season, the Impact have a mixed bag through the end of the year. On one hand they hold two games in hand over New York and one over Sporting but face a crucial Champions League match against Heredia on Tuesday night ahead of Saturday's game vs. the Fire.
Montreal still has a shot in the Champions League but not a great one… Will they turn their complete focus on the Fire and playoff race or try to double-dip this week?
4) Houston Dynamo
Current Points: 43 (12-10-7)
Games Remaining: 5 (3 home / 2 away)
Last 5 Games: WWLDL (7pts)
2013 Home/Away Record: 8-3-3 / 4-6-4
Maximum Possible Points: 58
Average Opponents Pts: 39.8
Remaining Schedule: 9/25 vs. WC**, 9/28 @ NE, 10/4 vs. MTL, 10/9 vs. SKC, 10/20 vs. NY, 10/24 @ AU**, 10/27 @ D.C.
Rundown: After going on an end of summer slump, Houston has run off two straight wins the past two weeks to move firmly into fourth place in the East. They play a crucial Champions League home match Wednesday night vs. W. Connection before opening up a four-game stretch all against teams in the Eastern Conference playoff picture. Three of those four matches are at home so after weathering a pretty difficult storm, its safe to say more than anyone, the Dynamo control their own fate in terms of being able to move up into one of the top three spots.
5) New England Revolution
Current Points: 40 (11-11-7)
Games Remaining: 5 (2 home / 3 away)
Last 5 Games: WLLDW (7pts)
2013 Home/Away Record: 7-5-3 / 4-6-4
Maximum Possible Points: 55
Average Opponents Pts: 43.0
Remaining Schedule: 9/28 vs. HOU, 10/5 @ NY, 10/12 @ MTL, 10/19 vs. CLB, 10/27 @ CLB
Rundown: One week New England is complaining about an offside goal being called back, the next they're benefiting from it and not saying a peep. This was the case on Diego Fagundez's tying goal in their 2-1 win over D.C. United. By virtue of that victory and the Fire's defeat at Columbus, the two sides traded places in the East race this weekend.
The good news for the Fire is that New England has the toughest remaining schedule in the East, with a huge showdown vs. streaking Houston on the horizon this weekend. Back-to-back trips to New York and Montreal follow that clash and could define the Revs season if they don't manage it the right way.
6) Chicago Fire
Current Points: 39 (11-12-6)
Games Remaining: 5 (2 home / 3 away)
Last 5 Games: LWDLD (5pts)
2013 Home / Away Record: 9-4-2 / 2-7-4
Maximum Possible Pts: 54
Average Opponents Pts: 34.8
Remaining Schedule: 9/28 vs. MTL, 10/4 @ D.C., 10/12 @ FCD, 10/19 vs. TFC, 10/27 @ NY
Rundown: The Fire lost a tough 3-0 result at Columbus on Saturday. It was a game the team went in with the mindset of taking three points and even after going down 1-0, they looked able to do it. Bakary Soumare's red card made that hope near impossible and as the Fire pushed forward they got caught out twice more.
They now welcome a struggling Montreal side to Toyota Park Saturday and should remember the situation they faced the Impact in back in August -- days after a disappointing U.S. Open Cup defeat to D.C. United, the Fire rebounded with a 2-1 win, a game that stands as one of the team's biggest results of the season.
While a 3-0 loss at this point in the season can be discouraging, perspective is important as the Fire remain just one point out of the top five. It is imperative for the team to take their remaining two home matches and find a way to get two results on the road to ensure they won't have to depend on other teams for a postseason berth.
7) Philadelphia Union
Current Points: 39 (10-10-9)
Games Remaining: 5 (2 home / 3 away)
Last 5 Games: LLDLD (2pts)
2013 Home/Away Record: 6-4-5 / 4-6-4
Maximum Possible Points: 54
Average Opponents Pts: 32.8
Remaining Schedule: 9/27 @ SKC, 10/5 vs. TFC, 10/12 @ D.C., 10/19 @ MTL, 10/26 vs. SKC
Rundown: The Union were idle this past weekend and will hope to have worked out the kinks that have him on the poorest form out of any team in the Eastern Conference playoff picture. Unfortunately for them, they face a huge match Friday night away to a Sporting side that can put themselves on the brink of a postseason berth with a win.
A loss in this match would be devastating to the team's chances but six points from their following two matches vs. Toronto and D.C. would certainly keep them afloat going into the final two weeks of the season.
8) Columbus Crew
Current Points: 35 (11-14-5)
Games Remaining: 4 (2 home / 2 away)
Last 5 Games: WWLWL (9pts)
2013 Home / Away Record: 7-5-3 / 4-9-2
Maximum Possible Points: 50
Average Opponents Pts: 42
Remaining Schedule: 9/29 @ FCD, 10/5 vs. SKC, 10/19 @ NE, 10/27 vs. NE
Rundown: The Crew continued their late-season run under interim head man Brian Bliss with a resounding 3-0 win over the Fire on Saturday. There's no doubt that the Crew need help from other teams but their attitude is in the right place to make a late-season run.
Every game is a must-win for the Crew from here on out and they'll shift their focus to a visit to FC Dallas where they haven't won since their MLS Cup championship season in 2008.


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After getting arrested, egging a house, and more, it's hard to imagine what these comedians will make fun of him for…
Last night, it was announced that the 20 year old, pop sensation heartthrob will have his own Comedy Central roast special. After 2014, which was pretty much the year of Bieber, it's a nice surprise to see Bieber being in on the joke and letting himself be made fun of on television by a bunch of professional comedians. Although the list of comedians partaking in such a momentous occasion has not been released, it is probably safe to say that this roast will be incredible.
This roast of Bieber will be following the roast of James Franco in 2013 and these comedians better step their game up for this A lister, no, AA lister. With his amazing track record of being arrested, egging his neighbor's house, peeing on a restaurant floor, partying it up recklessly in Miami, having assault charges, drag racing charges, photoshopped Calvin Klein underwear pictures, and much, much more, best wishes will go out to the comedians in this panel for trying to select which moment they make fun of. The roast is expected to air on March 7th, 2015 so be sure to check it out.
-Vinesh Vora


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Comfrey Root (Symphytum officinale)Latin Name: Symphytum officinale
Common Names: Comfrey Root, Slippery Root
Comfrey Root, with the Latin name of Symphytum officinale, is a perennial herb that features a long deep root, large rough leaves and bell-shaped flowers that range in color from cream to pink. The leaves of Symphytum officinale are similar in appearance to those found on the garden plant known as foxglove.
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Saint Laurent
Women's Star Print Scarf in Black
€495€198 (60%)
Size: One Size
- One Size Notify me
- Cashmere and silk blend in an ultra fine weave
- All-over star print
- Saint Laurent branding
- Frayed edges
- Squared silhouette
- Made in Italy
- Width: 135cm, Length: 135cm
Founded in 1961 by Yves Saint Laurent, the label is one of the most consistently celebrated French fashion brands to this day. Retaining its French irreverence and sleek craftsmanship, now with Anthony Vaccarello as creative director, the iconic label is pioneering a new era of 'forever classics'. With feminine allure teamed with decadent 1970s bohemia and psych rock, Saint Laurent offers a refined take on fashion with sharp tailoring, sheer blouses, babydoll dressses and slick leather jackets.
Vaccarello's first collection for SS17 is rooted in 80s influences and a youthful renegade aesthetic. Rejecting Parisian bourgeois, hard-cut leathers and sliced velvet silhouettes capture the brand's brash femininity; revealing windows of skin and fitted tight to the body. Vintage shapes are updated, reviving puffball-shouldered tops and dresses with deep sweetheart necklines and asymmetric elements. Low-slung boyfriend jeans and signature drainpipe skinnies are paired with sheer shirts, bustier tops and embellished cami tops whilst luxe basics are accompanied by patent jackets, lamé draped separates and contrast frilled pumps.
We offer Standard and Express Shipping
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*Excludes underwear, fragrance and cosmetic items which are non-returnable.
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As soon as we have shipped your order you will receive an email stating the tracking number. You can check the status of your order one to two hours after receiving the email via the link provided.
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Express - EUR 55 - 2/3 BD


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BelleBane and Bounty – SuicideGirls of the Day [PICTURES]
Meet BelleBane and Bounty, two SuicideGirls for the price of one. BelleBane is into time travel and cute panties and loves being a MILF. Bounty hates a person on a power trip, loves a good back rub and spends way too much time working. Let's hope we can pull these two ladies apart. On second thought, let's hope we get stuck in the middle while trying to pull these ladies apart.
Name: BelleBane
Into: Donnie Darko, time travel, aliens, kittens, cartoons, WW II, words, dreams, cute panties, laughter, being emo, obsessive love and random nonsense
Not into: Jerks, healing tattoos, scary clowns, hangovers, things that smell like popcorn, waiting long, roller coasters & lying
Makes me happy: Blueberry muffins
Makes me sad: Rain
Hobbies: Spending money that I don't have
5 things I can't live without:
- Eye Liner
- Fresh Ink
- A good lay
- Pizza
- Movies
Vices: Boys with tattoos, girls with tattoos, watching people get tattooed, convincing others to get tattooed, getting tattooed
Thoughts on SuicideGirls: I love it
I spend most of my free time: Being a MILF
- Mars Volta
- Deftones
- Incubus
- Linkin Park
- Red Hot Chilli Peppers
- Foo Fighters
- Paramore
- 30 Seconds to Mars
- Korn
- Muse
Name: Bounty
Location: USA
Into: Music, modeling, make-up, kisses, hugs, back rubs, soft skin, soft hair, secrets, sleeping, laughing, traveling, singing
Not into: People stuck on a power-trip, cocky attitudes, prejudgment, low self-esteem, ladies & gents with no self respect
Makes me happy: Accents, nice boys
Makes me sad: Being away from loved ones
Hobbies: Life
5 things I can't live without:
- YouTube – for musical purposes
- Wallet
- Contacts
- Face wash
- Gum
Thoughts on SuicideGirl: I love it
I spend most of my free time: Working! Fix that please
- For Today
- Bleed From Within
- Motionless In White
- Emmure
- A Day To Remember
- Switchfoot
- The Vitamin String
- Quartet
- Warhound
- We Drive Yeah
SuicideGirls is a community that celebrates alternative beauty and alternative culture from all over the world. Since 2001, tens of thousands of models have submitted millions of photos to their website hoping to become SuicideGirls.


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Griffin:Immunofluorescence Cell Staining
- 1 Immunofluorescence Cell Staining
- 2 Common fixatives to consider in protocol optimization
- 3 DAPI Counterstain
- 4 Antigen retrieval for cryostat tissue sections or cultured cells
Immunofluorescence Cell Staining
Below is a simple and clear approach to immunofluorescence. There are 3 common fixation methods listed below that are proven and effective for a multitude of adherent and suspension cells. 1% formalin PBS is a good starting fixation method. For a typical slide, it takes ~300 microliters to cover an area equal to 2 coverslips.
I) Culture cells to 50-70% confluency. Some cells (endothelial/epithelial) may require a higher level of confluency. For suspension cells in a regular petri dish or flask, transfer enough cells suspended in medium to each well of a poly-D-lysine-coated microslide. Then Incubate microslide in CO2 incubator from 1 hour to overnight to allow cells to attach to the bottom of the slide. The incubation time varies depending on the suspension cell used. Check for cell attachment using a microscope.
Gently aspirate medium from the wells, wash the attached cells briefly with 1x PBS and proceed with the Cell Staining Procedure of choice (i.e., protocol 6: Immunoperoxidase Cell Staining, or protocol 7: Immunofluorescence Cell Staining)
II) Fix the subconfluent cells appropriately, followed by 2 PBS washes (dipping the slides gently into separate beakers of PBS is one way to wash). Optional step: Antigen retireval (see below).
III) Blocking: Gently pipet 10% BSA, PBS onto the slide and incubate for 30 minutes rt.
IV) Primary Antibody Incubation: 1:50 in 2% serum/PBS for 60 minutes at room temperature followed by 3 separate and gentle PBS washes.
V) Secondary Antibody Incubation: 1:100 in 2% serum/PBS for 60 minutes at room temperature followed by 3 PBS washes and 1 dH2O wash.
At this point prop the slides at an angle to allow them to partially dry. (You will see liquid evaporate slowly off slide when faced up. Wait for all liquid to evaporate and then add the mounting medium) approximately 5 ul drop (no air bubbles) to each area (8 areas for 8 well plates) Gently drop the coverslip onto the slide and align. Allow it to settle. Place mounted slide in light proof box and place in 4 degree overnight priot to performing the visualization.
The mounting agent is two component, one powder and one liquid. When you add the liquid make sure all the powder is dissolved. The powder tends to be in chunks on the bottom of the the jar vial. Vortexing may not loosen up this chunk and into its dissolved state. Also let the soultion set for 5 minutes before pipeting to allow air bubbles to rise out of the glycerol. Molecular Probes Prolong Antifade Kit
Visualize under the appropriate fluorochrome filter.
Common fixatives to consider in protocol optimization
Methanol fixation (for cytoskeletal components)
The methanol fixation is an easy method; however, it frequently solubilizes and removes membrane bound antigens. By a simple precipitation of the protein, methanol only provides low structural preservation.
- Rinse the cover glass with PBS.
- Fix cells by incubating the cells in pre-cooled 100% methanol at -20 oC for 10 min.
- Wash cells with PBS.
Formaldehyde fixation (for membrane associated components)
Higher polymers, which are initially insoluble, are sold as a white powder known as paraformaldehyde.
- Rinse cells with PBS at room temperature.
- Fix in 3-4 % paraformaldehyde in PBS for 15 min at room temperature.
- Wash 3-times 5 min each with PBS containing 100 mM glycine.
- Permeabilize cells with 0.1% Triton X-100 in PBS for 1 to 4 min.
- Rinsed with PBS.
Formalin fixation (for membrane associated components)
The liquid known as formalin contains 37-40% of formaldehyde and 60-63% of water (by weight), with most of the formaldehyde existing as low polymers.
- Rinse cells with PBS at room temperature.
- Fix in 1 % Formalin in PBS (prepared from 10% buffered formalin PBS) for 10 min at room temperature.
- Wash 3-times with PBS
- Permeabilize cells with 0.1% Triton X-100 in PBS for 1 to 4 min.
- Rinsed with PBS.
DAPI Counterstain
4,6-Diamidino-2-phenylindole, 2HCl for DNA labeling. DAPI is a popular nuclear counterstain for use in multicolor fluorescent techniques. Its blue fluorescence stands out in vivid contrast to green, yellow or red fluorescent probes of other structures. DAPI stains nuclei specifically, with little or no cytoplasmic labeling. The following protocol is used for staining of tissue sections or for staining cultured cells on slides.
- Working concentration: 0.1 to 1 µg/mL DAPI.
- Staining Pattern: Nuclei will be stained bright blue.
- Suggested Use: Counterstain for immunofluorescence when green (FITC) or red (Texas Red) fluorescent marker is used.
- Excitation/Emission: ~359 nm/~461 nm
DAPI Counterstain Solution
DAPI Stock Solution (5mg/ml or 14.3 mM):
DAPI -------------------------------------------------------- 10 mg
Dimethylformamide (DMF) ------------------------------ 2 ml
Mix to dissolve; it may take some time to completely dissolve. Aliquot and store in –20 ºC.
DAPI Working Solution (100ng/ml or 300 nM in PBS):
DAPI stock solution --------------------------------------- 2 ul
PBS -------------------------------------------------------- 100 ml
Store this solution at 4ºC in brown bottle or wrapped with aluminum foil to protect from light. Incubate sections in dark for 30 minutes at room temperature.
Following reconstitution, aliquot and freeze (-20C) for long term storage or refrigerate (+4C) for short term storage. Stable for 1 years as supplied. Stock solutions stable for several weeks at +4C or for several months at -20C
Antigen retrieval for cryostat tissue sections or cultured cells
This is a simple method for antigen retrieval on aldehyde-fixed cryostat tissue sections or cultured cells. In many case, a brief 5 minutes pretreatment with 1% sodium dodecyl sulfate (SDS) produced a dramatic increase in staining intensity by immunohistochemistry and immunofluorescence.
Solutions and Reagents
1% Sodium Dodecyl Sulfate (SDS) in PBS:
- SDS ----------------------------- 1 g
- 0.01M PBS (pH 7.4) ---------------- 100 ml
Mix to dissolve.
- Rinse sections three times for 5 min each in PBS.
- Cover sections with 1% SDS solution and incubate for 5 minutes at room temperature.
- Rinse sections three times for 5 min each in PBS. It is important to wash sections well, otherwise residual SDS will denature the antibodies subsequently applied to sections.
- Incubate sections in serum blocking solution.
- Incubate in the primary antibody and complete immunohistochemical staining steps as desired.


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Shawn Zobel NFL Draft predictions and analysis are featured in numerous articles and publications online. Draft Headquarters is the #1 place for NFL Draft information!
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Adora Flow Bath Filler
Adora Flow Mini Basin Monobloc
Reference 13485 Code MBFW114P+ 6 In Stock
Order in next -1 days | 18 hrs | 46 mins
Adora Flow Mini Basin Monobloc.
The Adora Flow Mini Basin Monobloc features a stylish lever handle with curved spout that will transform any bathroom style. The mini design makes it a stunning option for the smaller bathroom.
Product Information:
- Minimum 0.2 bar pressure
- With click clack waste
- Projection: 114mm
- Height: 122mm
- Chrome finish
- Suitable for all plumbing systems
- Ideal for the smaller basin
- 5 Year Guarantee
- Matching bath tap options available
How Long will my delivery take?
Delivery times will vary depending on a number of factors including type of product ordered and whether it is an item that is carried in stock or a special order. Look out for the delivery times displayed on the product pages that act as a guide to delivery estimates.
Small items
include taps, shower systems, bathroom accessories and towel rails.
Medium items
include toilets, mirrors, basins and any other item not defined as
Large items
include toilets and basins, baths, shower cubicles, and any other item not defined as
£5.95 Delivery on this Item2-7 Days Estimated Delivery Time
Free Deliveryon All Orders over £500


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Carbon Steel 45 Degree Elbow, Press x Press Connection Type, 1-1/4" x 1-1/4" Tube Size
- Item # 36N790
- My Part #
- Mfr. Model # 25246
- Catalog Page # 2798
Using 360° Viewing:
- Rotate: Use top-to-bottom, side-to-side by use of mouse arrow.
- Zoom In: Double click on image.
- Zoom Out/Reset: Put photo at full zoom & then double click.
CAD Downloads
- Shipping Weight 1.05 lbs.
Product Details
Technical Specs
- Item 45 Degree Elbow
- Tube Fitting Material Carbon Steel
- Fitting Connection Type Press x Press
- Tube Size 1-1/4" x 1-1/4"
- Pipe Size - Pipe Fitting 1-1/4" x 1-1/4"
- Max. Pressure 125 psi
- Temp. Range -40 Degrees to 180 Degrees F
- Standards ANSI LC4/CSA 6.32: Fuel Gas Distribution Systems, IAPMO: Uniform Plumbing Code, ICC: International Mechanical Code, IFGC: International Fuel Gas Code, NFPA 54: National Fuel Gas Code, NFPA 58: Liquefied Petroleum Gas Code
- Compliance ABS: American Bureau of Shipping, CSA ANSI LC4/CSA 6.32, IAPMO ANSI LC4/CSA 6.32, ICC-ES PMG-1036, CSA B149.1, CRN: 0A14541.5 A/B
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Grainger is neither responsible for, nor does it endorse, the content of any product review or statement posted. Any statements posted constitute the statements of the poster and are not the statements of Grainger. The statements posted by Grainger employees with the Grainger employee badge represent the views of such employees and are not the statements of Grainger. Grainger makes no representations as to the appropriateness, accuracy, completeness, correctness, currentness, suitability, or validity of any product review or statements posted, including those posted by employees with the Grainger employee badge, and is not liable for any losses, injuries or damages which may result from any such product review or statements. Use of any linked web site provided in a product review or post is at the user's own risk.


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Sunday, January 1, 2012
The Better Team Won
The Giants were the better team. I really thought the Cowboys were better than this but I guess I was just seeing things with my heart and not my head.
The Cowboys secondary is terrible. It is time for Newman to retire or get cut. Why don't the Cowboys DB's ever make a play on the ball? Eli Manning basically closed his eyes and threw the ball up to Victor Cruz with the Cowboys down 7 trying to get the Giants off the field on 3rd down. Scandrick had position but just got outplayed by Cruz once the ball was in the air.
The defense and the offensive line lost the game. A 21-0 deficit was just too much to overcome without a few boneheaded mistakes by the Giants. Maybe with a full offseason the defense will be able to grasp the complex Rob Ryan defense. There was way too much confusion pre-snap in the defense.
I knew the offensive line would be an issue heading into the season when the Cowboys cut 3 former pro bowlers. Unfortunately their youth and inexperience really showed in the most important game of the season.
Tony Romo is a great quarterback. He was the only reason the game wasn't a complete disaster in the 2nd half. He was sacked 6 times and I think there could have been at least 6 more if he wasn't making free rushers miss.
Why did it take until halftime for the Cowboys to adjust on offense and defense? By halftime it was really too late to comeback without playing nearly flawlessly in the 2nd half.
I came to the conclusion after the first embarrassing loss to the Eagles that the Cowboys were a mediocre team. So 8-8 is a fitting finish to the season.
There is a lot of work to do in the offseason. The secondary needs a serious makeover. And the offensive line needs major improvement.
5 comments:
- AnonymousJanuary 1, 2012 at 11:04 PM
Rob Ryan needs to go and if J.G. is to stay, he needs to be regulated as a head coach and not an O.C. I've made the latter point now for over 2 years and I will not back off of it. Amazing what could've happened if we kept Sean Payton. No use in crying over spilled milk...ReplyDelete
The middle of the defense needs a big shift. Ratliff @ end and finally get a real 3-4 nose. Lissemore quietly had a great season for the amount of snaps he got. Hopefully Carter steps in well next year.
Brookings, James, Spencer, Newman, and Elam (hopefully) will be gone. I wholeheartedly believe we do not necessarily need a top-5 defense (although it would be nice) but play-makers that take advantage of opportunities.
OLB, ILB, Nose, G, S, and CB are our needs. Silver lining?? pick 14 and we play 3 seeds next year with an already difficult schedule. I'm getting tired of looking ahead to next year.
There is one interesting thing I hope will be discussed. There will be a lot of rumors about H.C. and new Coordinators etc. I'm interested to hear if J.J. will reconsider bringing in Dan Reeves or someone of their caliber to evaluate talent...just a random thought.
- AnonymousJanuary 2, 2012 at 12:38 AM
I think that this season was more difficult because the Cowboys kept giving us hope. You knew that they weren't a good team and that they'd get absolutely crushed by either the Packers or the Saints but they kept sticking around. It's just been so long since they won anything. One playoff win in the last 15 years for America's Team is beyond pathetic. Next year Romo will be another year older. He shows signs of greatness but he has for years now. They had a chance for the playoffs, failed in every way imaginable and ended the season 8-8 and in third place in the division, technically behind even the Eagles. Another wasted season.ReplyDelete
- AnonymousJanuary 2, 2012 at 7:56 AM
We all need to realize after so many years of this....17 to be exact.....only stop attending games will slow down Jerry's idiotic decision making.ReplyDelete
- AnonymousJanuary 3, 2012 at 9:28 PM
Love your blog man. Im tired of ignorant Cowboy fans. Glad you speak the truth.ReplyDelete
- AnonymousJanuary 8, 2012 at 12:34 PM
The Cowboys have the worst secondary in the league, the absolute worse. The Newman experiment has been a failure. Jenkins is not much better. Elam, Scandrick, Sensabaugh and whatever other tomato-can corner they found in a Dallas nightclub need to go. I'm not a huge fan of the SEC, but they do produce secondary guys like clockwork. Go out and draft as many of them as possible, and hope that some of them work out.ReplyDelete


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A Practical and Biblical Approach to Witnessing
Response evangelism is possibe because God takes the initiative in salvation.
Every believer is called to be a witness for our Lord. But research shows that in any given church almost all the personal evangelism is done by less than 10 percent of the congregation.read more
Response evangelism is possibe because God takes the initiative in salvation.
Times of great crisis provide timely opportunities to share our faith with non-Christians.
Whenever possible, encourage non-Christians to read the life of Christ. I recommend giving them the Gospel of Mark.
Why don't more Christians tell others about Jesus Christ? For most people the real problem is lack of confidence.
We were created for a reason - a divine purpose. If we clearly share the truth about Jesus, even briefly, we should trust the Holy Spirit to bear witness to that truth in the mind and heart of a person.
God has a plan for each of our lives. Two great moments in our lives are when we are born and when we discover why we were born.
Jesus directed His disciples' focus to the task before them of being witnesses to a lost world in the power of the Spirit.
The apostle Peter's preaching in the Book of Acts always answers two basic questions: Who was Jesus? and Why did He give His life?
What does somebody really need to know to believe in Jesus and receive his gift of forgiveness and eternal life? In the book of Acts, Peter's preaching always focused on two things: Jesus' identity, and the purpose of his mission on earth.


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To access the New Chat Room, click on to the icon to your left.
The Mets have more than a few pitching questions heading into next year, and Mike Pelfrey is one of them. After a brilliant start, Pelfrey hit the skids in July, seemed to right himself, then was hit hard again. He's coming off a good start and the Mets want him to finish on a roll to give him the confidence heading into the offseason.
With Johan Santana injured, unless the Mets make a stunning move in the offseason, Pelfrey will go to spring training as the No. 1 starter.
The Mets will be seeking their first four-game sweep of a team tonight at home against Pittsburgh.


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Young woman working on a computer. Tunis, Tunisia. Photo: Arne Hoel / World Bank
"Girls programming isn't just a cool thing; it's also doubling the chances of developing innovative tools and making the world a better place for everyone." These words are from my friend Julie, who has been working as a web developer for the last four years. She has also been involved in a few volunteer programs in Africa, mainly to train young women on IT tools.
If you are able to read this blog, it means you know how to use a computer or another digital device, such as a smartphone. And the great thing is that you may be a boy or a girl, from a rich or a poor region of the world. You could be anyone, really. Indeed, the digital revolution is not only happening in the richest countries. The generation of the "Digital Natives" is spreading all over the world, from South Asia to Africa, from Russia to the Caribbean and so on. And this might be the greatest opportunity for the world to close the gender gap.
Not everybody is as lucky as I was to be a girl born in France, with free access to computers and Internet in school. In many regions of the world girls suffer from having no (or little) access to education and jobs. Nevertheless, things are changing for the better. Globally, the gender education gap has been gradually shrinking since 1999 and today the number of girls and women who are passionate about new technologies is growing at an astonishing pace, especially in Africa.
As Thuli Sibeko, organizer of the annual Girls in ICT event in South Africa, recently explained in an interview with IT News Africa, the technology sector offers "an extraordinary opportunity" for girls and young women to forge successful careers. She also warned that governments need to actively encourage young girls to consider technology as a career option in order to address skills shortages and unemployment
On October 2012, the United Nations agency for information & communication technologies (ITU) launched the Tech Needs Girls initiative to "inspire more girls to embrace technology and invent the future" as well as a multilingual portal to help girls and young women prepare for and pursue a technology career.
The world is just realizing that the words "girls" and "technology" go along with inventing a better world. What do you think? Tell us in the comments.
Truly the access of
Truly the access of technology ( internet,etc) to girls can change the world for better as we all know that girls actually run a home so in a sense a community , in a sense the whole nation. The underlying problem is related with the developing and emerging countries where we need to provide the girls/women the access of technology/internet so that they can pave a way for them and for their next generation. This has the potential to unleash a fission reaction which would result in a knowledge society and eventually help a nation with a skilled population and contributing towards the nation's success.
No doubt Technology is having
No doubt Technology is having a large impact on girls & women. They are now much aware about their rights & strengths through TV/Mobile/Computers. Growth of technology has created opportunities also in technology sector, definitely for women also. The best thing is that with introduction of technology Men are also recognizing the rights of women.