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Also for CCNB1 (NM_031966)
|This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids 120-131 of Human Cyclin B1 protein.|
|human, rat, dog and chimpanzee, mouse, hamster.
||ELISA: 1:50,000, WB: 1:100 - 1:1,000, IP: 1:100
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|Cyclin B1 (also called CCNB1 and G2/mitotic-specific cyclin B1) is a regulatory protein involved in mitosis. The gene product complexes with p34(cdc2) to form a serine/threonine kinase holoenzyme complex called the maturation-promoting factor (MPF). Two alternative transcripts have been found a constitutively expressed transcript and a cell cycle-regulated transcript that is expressed predominantly during G2/M phase. The different transcripts result from the use of alternate transcription initiation sites. S126 phosphorylation, the site of phosphorylation recognized by this antibody, is involved in an auto-phosphorylation of the cdk -cyclin complex. Recognition of this phosphorylated site suggests the antibody is useful to detect the active form of the complex and may serve as a marker of mitosis.
|Homo sapiens cyclin B1 (CCNB1)|
Hedgehog signaling pathway
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Western blot using Affinity Purified anti-Cyclin B<sub>1</sub> pS126 antibody shows detection of a band ~48 kDa corresponding to phosphorylated human Cyclin B<sub>1</sub> (arrowheads) in various whole cell lysates. Lysates tested were lane 1 - Hela (cervical carcinoma), lane 2 - H23 (lung carcinoma), lane 3 - Hep3b (Hepatocarcinoma), lane 4 - T98G (Glioblastoma) and lane 5 - Daudi (B cell lymphoblast). 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 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 1:5,000 dilution of HRP conjugated Gt-a-Rabbit IgG [H&L] MX (611-103-122) 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. Personnel Communication, Luca Cote, Temple U.