05-373
Anti-Cyclin B1 Antibody, clone GNS3 (8A5D12)
clone GNS3 (8A5D12), Upstate®, from mouse
Synonym(s):
G2/mitotic-specific cyclin B1, cyclin B1
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
mouse
Quality Level
antibody form
purified antibody
antibody product type
primary antibodies
clone
GNS3 (8A5D12), monoclonal
species reactivity
human, mouse
packaging
antibody small pack of 25 μg
manufacturer/tradename
Upstate®
technique(s)
immunohistochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgG
NCBI accession no.
UniProt accession no.
shipped in
dry ice
target post-translational modification
unmodified
Gene Information
human ... CCNB1(891)
mouse ... Ccnb1(268697)
General description
G2/mitotic-specific cyclin-B1 (UniProt: P14635; also known as Cyclin B1) is encoded by the CCNB1 (also known as CCNB) gene (Gene ID: 891) in human. Cyclins are the regulatory subunits of the cell cycle-dependent kinases (CDKs) that are responsible for the phosphorylation of several cellular targets. Cyclins contain the nuclear localization sequence (NLS) that help move CDKs into the nucleus. They also contain PEST (Pro, Glu, Ser, and Thr) sequences that target them for degradation by the ubiquitin-proteasomal pathway. Once the CDKs have completed their role, they undergo a rapid programmed proteolysis via ubiquitin-mediated delivery to the proteasome complex. Cyclin B1, a regulatory protein involved in mitosis, complexes with CDK1 to form the maturation-promoting factor (MPF). It is shown to be essential for the control of the cell cycle at the G2/M (mitosis) transition. It accumulates steadily during G2 phase and is abruptly destroyed at mitosis. Hence, the cyclin B1-CDK1 complex is considered to be a key regulator for mitotic entry. This complex phosphorylates a number of proteins prior to mitotic entry. Although five serine phosphorylation sites are described for cyclin B1, (Ser 116, 126, 128, 133, and 147), serine 133 phosphorylation by PLK1 regulates the entry of Cyclin B1-CDK1 complex into the nucleus during prophase. At the end of mitosis, cyclin B1 is rapidly removed by a ubiquitin ligase (anaphase-promoting complex/cyclosome) loaded with the targeting subunit CDC20. Activated cyclin B1-CDK1 complex is reported to catalyze its own destruction by stimulating the activity of APC. (Ref.: Van Zon, W., et al. (2010). J. Cell. Biol. 190(4); 587-602; Yuan, J., et al. (2004). Oncogene 23(34); 5843-5852).
Specificity
In addition to human, weak species cross-reactivity was observed with mouse.
This antibody is specific for human cyclin B1, Mr 58 kDa. Does not cross-react with cyclin A or cyclin D.
Application
Immunoprecipitation:
2 μg of a previous lot immunoprecipitated human cyclin B1 and cdc2 kinase from 500 μg of A431 RIPA lysate as determined by a subsequent immunoblot of the immunoprecipitate using anti-cdk1/cdc2 (PSTAIR), (Catalog # 06-923).
Western Blotting Analysis: 0.1 mg/mL of this antibody detected Cyclin B1 in A431 cell lysate.
Immunohistochemistry (Paraffin) Analysis: A 1:250 and 1:50 dilutions of this antibody detected Cyclin B1 in Human tonsil and Human bone marrow tissue sections, respectively.
2 μg of a previous lot immunoprecipitated human cyclin B1 and cdc2 kinase from 500 μg of A431 RIPA lysate as determined by a subsequent immunoblot of the immunoprecipitate using anti-cdk1/cdc2 (PSTAIR), (Catalog # 06-923).
Western Blotting Analysis: 0.1 mg/mL of this antibody detected Cyclin B1 in A431 cell lysate.
Immunohistochemistry (Paraffin) Analysis: A 1:250 and 1:50 dilutions of this antibody detected Cyclin B1 in Human tonsil and Human bone marrow tissue sections, respectively.
Quality
Routinely evaluated by western blot analysis on RIPA lysate from human A431 carcinoma cells.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected cyclin B1 in RIPA lysates from human A431 carcinoma cells.
Western Blot Analysis:
0.5-2 μg/mL of this lot detected cyclin B1 in RIPA lysates from human A431 carcinoma cells.
Target description
58 kDa
Linkage
Replaces: 04-220
Physical form
Format: Purified
Protein G purified mouse IgGs in storage buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide. Frozen at -20°C.
Analysis Note
Control
Positive Antigen Control: Catalog #12-301, non-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for minigels.
Positive Antigen Control: Catalog #12-301, non-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for minigels.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
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May, M; Schelle, I; Brakebusch, C; Rottner, K; Genth, H
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Kalyan Dulla et al.
Molecular & cellular proteomics : MCP, 9(6), 1167-1181 (2010-01-26)
Reversible protein phosphorylation is a key regulatory mechanism of mitotic progression. Importantly, protein kinases themselves are also regulated by phosphorylation-dephosphorylation processes; hence, phosphorylation dynamics of kinases hold a wealth of information about phosphorylation networks. Here, we investigated the site-specific phosphorylation
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