CCNB1 (phospho S126) polyclonal antibody
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More Files
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Western Blot (Cell lysate)
Western blot using CCNB1 (phospho S126) polyclonal antibody (Cat # PAB10042) shows detection of a band ~48 kDa corresponding to phosphorylated human CCNB1 (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 ug 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 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
Personnel Communication, Luca Cote, Temple U.
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic phosphopeptide of CCNB1.
Immunogen
Synthetic phosphopeptide corresponding to residues surrounding S126 of human CCNB1.
Host
Rabbit
Reactivity
Chimpanzee, Dog, Human, Rat
Specificity
This antibody is specific to phosphorylated human Cyclin B1 protein at the pS126 residue. Minimal reactivity is expected with the non-phosphorylated form of the protein.
Form
Liquid
Quality Control Testing
Antibody Reactive Against Synthetic Peptide.
Recommend Usage
ELISA (1:50000)
Western Blot (1:100-1:1000)
The optimal working dilution should be determined by the end user.Storage Buffer
In 20 mM KH2PO4, 150 mM NaCl, pH 7.2 (0.01% sodium azide)
Storage Instruction
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
Western Blot (Cell lysate)
Western blot using CCNB1 (phospho S126) polyclonal antibody (Cat # PAB10042) shows detection of a band ~48 kDa corresponding to phosphorylated human CCNB1 (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 ug 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 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
Personnel Communication, Luca Cote, Temple U.Immunoprecipitation
Enzyme-linked Immunoabsorbent Assay
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Gene Info — CCNB1
Entrez GeneID
891Protein Accession#
P14635;NP_114172Gene Name
CCNB1
Gene Alias
CCNB
Gene Description
cyclin B1
Omim ID
123836Gene Ontology
HyperlinkGene Summary
The protein encoded by this gene is a regulatory protein involved in mitosis. The gene product complexes with p34(cdc2) to form 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. [provided by RefSeq
Other Designations
G2/mitotic-specific cyclin B1
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Interactome
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Pathway
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Disease
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Publication Reference
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Human papillomavirus type 16 E1 E4-induced G2 arrest is associated with cytoplasmic retention of active Cdk1/cyclin B1 complexes.
Davy CE, Jackson DJ, Raj K, Peh WL, Southern SA, Das P, Sorathia R, Laskey P, Middleton K, Nakahara T, Wang Q, Masterson PJ, Lambert PF, Cuthill S, Millar JB, Doorbar J.
Journal of Virology 2005 Apr; 79(7):3998.
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p53 is a NF-Y- and p21-independent, Sp1-dependent repressor of cyclin B1 transcription.
Innocente SA, Lee JM.
FEBS Letters 2005 Feb; 579(5):1001.
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Regulation of activation-induced Fas (CD95/Apo-1) ligand expression in T cells by the cyclin B1/Cdk1 complex.
Torgler R, Jakob S, Ontsouka E, Nachbur U, Mueller C, Green DR, Brunner T.
The Journal of Biological Chemistry 2004 Sep; 279(36):37334.
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Human papillomavirus type 16 E1 E4-induced G2 arrest is associated with cytoplasmic retention of active Cdk1/cyclin B1 complexes.
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