![EEF2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit monoclonal to EEF2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab75748.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 95kDa; EEF2](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/11883_EEF2-Primary-antibodies-ab75748-3.jpg)
EEF2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit monoclonal to EEF2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab75748.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 95kDa; EEF2
![All lanes : Anti-EEF2 antibody [EP880Y] (ab75748) at 1/10000 dilutionLane 1 : 293 cell lysateLane 2 : HeLa cell lysateLane 3 : A431 cell lysateLane 4 : NIH3T3 cell lysateLysates/proteins at 10 µg per lane.SecondaryHRP labelled goat anti-rabbit at 1/2000 dilution](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/11884_ab75748_1.jpg)
All lanes : Anti-EEF2 antibody [EP880Y] (ab75748) at 1/10000 dilutionLane 1 : 293 cell lysateLane 2 : HeLa cell lysateLane 3 : A431 cell lysateLane 4 : NIH3T3 cell lysateLysates/proteins at 10 µg per lane.SecondaryHRP labelled goat anti-rabbit at 1/2000 dilution
ab75748 at 1/100 dilution staining EEF2 in human kidney carcinoma tissue.
ICC/IF image of ab75748 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75748, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Overlay histogram showing HeLa cells stained with ab75748 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75748, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.