RECK Antibody / Reversion-inducing cysteine-rich protein with Kazal motifs (FY13235)

Flow Cytometry analysis of human JK cells using anti-RECK antibody. Overlay histogram showing JK cells stained with (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-RECK 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 RECK using anti-RECK antibody. Lane 1: human U251 whole cell lysates, Lane 2: human Jurkat whole cell lysates, Lane 3: human SH-SY5Y whole cell lysates, Lane 4: rat C6 whole cell lysates, Lane 5: mouse LLC 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-RECK 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 detection of RECK shows a single band migrating just below 100 kDa across multiple lysates. Although the calculated mass is ~106 kDa, RECK frequently runs between ~95 and 115 kDa due to variable glycosylation and maturation of this GPI-anchored protein.