Western blot analysis of P-Selectin/SELP using anti-P-Selectin/SELP antibody (M01241).
Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: human HEL whole cell lysates,
Lane 3: human THP-1 whole cell lysates,
Lane 4: human SH-SY5Y whole cell lysates,
Lane 5: human K562 whole cell lysates,
Lane 6: human Jurkat whole cell lysates,
Lane 7: human SIHA whole cell lysates,
Lane 8: human RT4 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-P-Selectin/SELP antigen affinity purified monoclonal antibody (M01241) at 1: 500 overnight at 4°C, 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 is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for P-Selectin/SELP at approximately 91 kDa. The expected band size for P-Selectin/SELP is at 91 kDa.