AXIN1 Antibody / Axin 1 (FY13148)

Immunofluorescent staining of AXIN1 using anti-AXIN1 antibody (green) and anti-Beta Tubulin antibody (red). AXIN1 was detected in immunocytochemical section of HELA cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-AXIN1 antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. Cy3 Conjugated Goat Anti-Rabbit IgG and DyLight 488 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of AXIN1 using anti-AXIN1 antibody. Lane 1: human HT-1080 whole cell lysates, Lane 2: human whole cell lysates, Lane 3: human K562 whole cell lysates, Lane 4: mouse brain tissue lysates, Lane 5: mouse thymus tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-AXIN1 antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. AXIN1 antibody detects a predominant band just above 100 kDa with weaker bands above and below across the indicated samples. Although the predicted mass is ~96 kDa, AXIN1 frequently migrates slower due to multisite phosphorylation, and additional higher and lower species likely reflect ubiquitinated forms and limited proteolysis.