Immunohistochemical analysis of paraffin embedded
breast carcinoma, using primary CDK2
(Phospho-Thr160) ab28808 at 1:100 dilution and Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L) secondary antibody. Left:
Untreated; Right: Treated with synthesized
Western blot - GSK3 alpha (phospho S21) antibody (ab28808)
All lanes : Anti-GSK3 alpha (phospho S21) antibody (ab28808) at 1/1000 dilution
Lane 1 : Extracts from ovary cancer
cells. Lane 2 : Extracts from ovary cancer
cells. with synthesised non phosphopeptide at 1 µg/ml Lane 3 : Extracts from ovary cancer
cells. with synthesised phosphopeptide at 1 µg/ml
Lysates/proteins at 30 µg per lane.
Predicted band size : 50 kDa Observed band size : 50 kDa
ICC/IF image of ab28808 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 (ab28808, 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.
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