CCNB1 Antibody

Western blot using Biorbyt's Affinity Purified anti-Cyclin B1 pS126 antibody shows detection of a band ~48 kDa corresponding to phosphorylated human Cyclin B1 (arrowheads) in various whole cell lysates. Lane 1: Hela (cervical carcinoma) [p/n orb348668], Lane 2: H23 (lung carcinoma), Lane 3: Hep3b (Hepatocarcinoma), Lane 4: T98G (Glioblastoma), and Lane 5: Daudi (B cell lymphoblast) [orb692723]. Each lane contains approximately 50 µg of lysates, separated by 12% SDS-PAGE using a 5% stack run at 100 volts until the dye front cleared the bottom of the gel. Transfer occurred overnight at 4°C at 15 mAmps. The membrane was blocked with 5% non-fat dry milk in TTBS (p/n orb348624) for 1 h at room temperature followed by addition of a 1:100 dilution of the antibody allowed to react for 2h at room temperature. After washes with TTBS a 1:5000 dilution of HRP conjugated Gt-a-Rabbit IgG [H&L] MX (p/n orb347654) was added for 1 h at room temperature. After additional washes the membrane was incubated with ECL mix 1:1 for ~3 min. Excess detection solution was drained off and the membrane was exposed to Kodak film X-omat blue XB-1 for about 20 sec. Other detection systems will yield similar results.