OSTM1 Antibody / Osteopetrosis-associated transmembrane protein 1 (FY12777)

Immunofluorescent staining of OSTM1 using anti-OSTM1 antibody (green) and anti-Beta Tubulin antibody (red). OSTM1 was detected in an immunocytochemical section of human SIHA cells. 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-OSTM1 antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG and Cy3 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 OSTM1 using anti-OSTM1 antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human Raji whole cell lysates, Lane 2: human U251 whole cell lysates, Lane 3: human whole cell 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-OSTM1 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 an ECL Plus Western Blotting Substrate. A strong band is detected near the expected ~37 kDa size, corresponding to the mature glycosylated form of OSTM1, along with a faint lower band around 35 kDa likely representing a partially processed species.