Product Pathways - Cell Cycle / Checkpoint
Cell Cycle Regulation Antibody Sampler Kit II #9870
|9870S||1 Kit (8 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Phospho-cdc2 (Tyr15) Antibody #9111||40 µl||W, IP||H, M, R, Mk, Dm, X, Sc||34||Rabbit|
|Cyclin A2 (BF683) Mouse mAb #4656||40 µl||W, F||H||55||Mouse IgE|
|Cyclin B1 Antibody #4138||40 µl||W, IF-IC||H, M, R, Hm, Mk||58||Rabbit|
|Cyclin E2 Antibody #4132||40 µl||W||H||48||Rabbit|
|Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377||40 µl||W, IF-IC, F||H, M, R, Mk, Z||17||Rabbit IgG|
|Myt1 Antibody #4282||40 µl||W, IHC-P||H, M, R||X||60 to 70||Rabbit|
|p21 Waf1/Cip1 (12D1) Rabbit mAb #2947||40 µl||W, IP, IHC-P, IF-IC, F||H, Mk||Dg||21||Rabbit IgG|
|Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb #4910||40 µl||W, IP||H||M, R, Mk, X, Z, B||95||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|Anti-mouse IgG, HRP-linked Antibody #7076||100 µl||Horse|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Dm=D. melanogaster, X=Xenopus, Sc=S. cerevisiae, Hm=Hamster, Z=Zebrafish
Western blot analysis of extracts from various cell types using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947.
Western blot analysis of extracts from HeLa cells, untreated or treated with nocodazole (0.1 mg/ml for 18 hours), using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377 (upper) or Histone H3 Antibody #9715 (lower). Phospho-specificity of the antibody is shown by further treatment of the lysate with λ phosphatase.
Western blot analysis of extracts from MCF7, SK-N-MC and HeLa cells, untreated or treated with the proteasome inhibitor MG-132, using Cyclin E2 Antibody #4132.
Western blot analysis of extracts from various cell types using Cyclin B1 Antibody #4138.
Western blot analysis of extracts from HT-29 cells, untreated (lane 1), nocodazole-treated (50 ng/ml, lane 2), or UV-treated (lane 3), using Myt1 Antibody #4282.
Western analysis of extracts from HeLa cells, untreated or treated with doxorubicin (0.5 μM, 24 hours) or with nocodazole (50 ng/ml, 24 hours), using Cyclin A (BF683) Mouse mAb #4656.
Western blot analysis of extracts from A431 and H441 cells, untreated or EGF-treated, using Phospho-Wee1 (Ser642) (D47G5) Rabbit mAb #4910 (upper) or Wee1 Antibody #4936 (lower).
The Cell Cycle Regulation Sampler Kit II provides an economical means of evaluating cell cycle proteins. The kit contains enough primary and secondary antibodies to perform four western blot experiments.
Specificity / Sensitivity
Each antibody in the Cell Cycle Regulation Sampler Kit II detects endogenous levels of its target protein and does not typically cross react with other family members. Activation state antibodies recognize target proteins only when phosphorylated at the indicated residue. Phospho-cdc2 (Tyr15) Antibody detects endogenous levels of cdc2, CDK2 and CDK5 when phosphorylated at Tyr15 and yeast orthologue of cdc2 (cdc28) when phosphorylated at Tyr19.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues near the amino terminus of human cyclin B1, the carboxy-terminus of human cyclin E2, or the middle of mouse and human Myt1. Activation state polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr15 of human cdc2. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues near the carboxy-terminus of human p21 or recombinant human cyclin A2 protein. Activation state monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser10 of human histone H3 or residues surrounding Ser642 of human Wee1.
The critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Tyr15 and Thr14 (1). Phosphorylation of cdc2 by the protein kinases Wee1 and Myt1 at Thr14 and Tyr15 results in inhibition of cdc2 (2,3). Progression through the G1/S checkpoint and initiation of DNA replication requires cyclin E; traversing the G2/M checkpoint to initiate mitosis requires cyclin B, and cyclin A may be required for both S-phase and M-phase (4). The versatile p21 cyclin-dependent kinase inhibitor, which interacts with several cyclin-CDK complexes to regulate cyclin-CDK during the cell cycle, is regulated by phosphorylation and ubiquitin-mediated degradation (5). Phosphorylation of histone H3 at Ser10 and neighboring residues correlates with chromosomal condensation, which is essential for segregation of chromosomes during mitosis (6).
- Norbury, C. et al. (1991) EMBO J 10, 3321-9.
- McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85.
- Wells, N.J. et al. (1999) J Cell Sci 112 ( Pt 19), 3361-71.
- Pagano, M. et al. (1992) EMBO J 11, 961-71.
- Abbas, T. and Dutta, A. (2009) Nat Rev Cancer 9, 400-14.
- Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.