COLGALT1 Antibody / Collagen beta(1-O)galactosyltransferase 1 (FY12656)

Flow Cytometry analysis of HEL cells using anti-COLGALT1 antibody. Overlay histogram showing HEL cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-COLGALT1 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Western blot analysis of GLT25D1/COLGALT1 using anti-COLGALT1 antibody. Lane 1: human Hela whole cell lysates, Lane 2: human SH-SY5Y whole cell lysates, Lane 3: human THP-1 whole cell lysates, Lane 4: human U251 whole cell lysates, Lane 5: rat C6 whole cell lysates, Lane 6: mouse NIH/3T3 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-COLGALT1 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. Western blot analysis of COLGALT1 using anti-COLGALT1 antibody. A major doublet is observed at ~80-85 kDa in human samples, consistent with reported glycosylated forms of the protein, while mouse and rat lysates show a single band near the predicted 72 kDa corresponding to the less glycosylated ortholog.