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All lanes : Anti-AATF antibody (ab65248) at 1/500 dilution
Lane 1 : HeLa cell extract
Lane 2 : HepG2 cell extract
Lane 3 : COLO205 cell extract
Lane 4 : HUVEC cell extract
Lane 5 : HUVEC cell extract with immunising peptide at 5 µg
Lysates/proteins at 5 µg per lane.
Predicted band size : 62 kDa
Observed band size : 62 kDa
ICC/IF image of ab65248 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab65248, 1µg/ml) overnight at +4ºC. 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.
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