Product Pathways - Cell Cycle / Checkpoint

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Cell Cycle Regulation Antibody Sampler Kit II #9870

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Isotype
Phospho-cdc2 (Tyr15) Antibody #9111	40 µl	W IP	H M R Mk Dm X	34	Rabbit
Cyclin A2 (BF683) Mouse mAb #4656	40 µl	W F	H	55	Mouse IgG2b
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

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  Dm=D. melanogaster  X=Xenopus  Z=Zebrafish  B=Bovine  Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

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.

Description

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.

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.

Background

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).

1. Norbury, C. et al. (1991) EMBO J 10, 3321-9.
2. McGowan, C.H. and Russell, P. (1993) EMBO J 12, 75-85.
3. Wells, N.J. et al. (1999) J Cell Sci 112 ( Pt 19), 3361-71.
4. Pagano, M. et al. (1992) EMBO J 11, 961-71.
5. Abbas, T. and Dutta, A. (2009) Nat Rev Cancer 9, 400-14.
6. Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

2947:

Flow, IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

3377:

Flow, Immunofluorescence, Western Blotting

4132:

Western Blotting

4138:

Immunofluorescence, Western Blotting

4282:

IHC / Paraffin, Western Blotting

4656:

Flow, Western Blotting

4910:

Immunoprecipitation, Western Blotting

9111:

ELISA Peptide, Immunoprecipitation, Western Blotting

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and 7,429,487) from Epitomics, Inc.

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For Research Use Only. Not For Use In Diagnostic Procedures.