Product Pathways - Cell Cycle / Checkpoint

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Cell Cycle/Checkpoint Antibody Sampler Kit #9917

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
Phospho-Chk1 (Ser345) (133D3) Rabbit mAb #2348	40 µl	W IF-IC F	H M R Mk	56	Rabbit IgG
Phospho-Chk2 (Thr68) Antibody #2661	40 µl	W IP IF-IC F	H Mk	62	Rabbit
Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb #8516	40 µl	W IP IHC-P IF-IC F	H M R Mk	110	Rabbit IgG
Phospho-Rb (Ser795) Antibody #9301	40 µl	W IP	H R Mk	110	Rabbit
Phospho-p53 (Ser15) Antibody #9284	40 µl	W IP IF-IC ChIP	H M R Mk (Mi) (B) (Pg)	53	Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074	100 µl				Goat

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

Specificity / Sensitivity

Phospho-cdc2 (Tyr15) Antibody detects cdc2 and cdk2 only when phosphorylated at Tyr15 and does not cross-react with cdk4, cdk6 or cdk7. Phospho-Chk2 (Thr68) Antibody detects Chk2 only when phosphorylated at Thr68 and does not react with nonphosphorylated Chk2. Phospho-Chk1 (Ser345) Antibody detects Chk1 only when phosphorylated at Ser345 and does not cross-react with other proteins. Phospho-Rb (Ser795) and (Ser807/811) Antibodies detect Rb only when phosphorylated at the indicated sites and do not cross-react with Rb phosphorylated at other sites. Phospho-p53 (Ser15) Antibody detects p53 only when phosphorylated at Ser15 and does not react with nonphosphorylated p53.

Description

The Cell Cycle/Checkpoint Antibody Sampler Kit provides a fast and economical means of evaluating multiple proteins involved in the cell cyle and checkpoint control. The kit contains enough primary and secondary antibody to perform four Western blot experiments.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr15 of human cdc2; Thr68 of human Chk2; Ser795 and Ser15 of human p53. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser807/811 of human Rb protein and residues surrounding Ser345 of human Chk1.

Background

The cell division cycle demands accuracy to avoid the accumulation of genetic damage. This process is controlled by molecular circuits called "checkpoints" that are common to all eukaryotic cells (1). Checkpoints monitor DNA integrity and cell growth prior to replication and division at the G1/S and G2/M transitions, respectively. The cdc2-cyclin B kinase is pivotal in regulating the G2/M transition (2,3). Cdc2 is phosphorylated at Thr14 and Tyr15 during G2-phase by the kinases Wee1 and Myt1, rendering it inactive. The tumor suppressor protein retinoblastoma (Rb) controls progression through the late G1 restriction point (R) and is a major regulator of the G1/S transition (4). During early and mid G1-phase, Rb binds to and represses the transcription factor E2F (5). The phosphorylation of Rb late in G1-phase by CDKs induces Rb to dissociate from E2F, permitting the transcription of S-phase-promoting genes. In vitro, Rb can be phosphorylated at multiple sites by cdc2, cdk2, and cdk4/6 (6-8). DNA damage triggers both the G2/M and the G1/S checkpoints. DNA damage activates the DNA-PK/ATM/ATR kinases, which phosphorylate Chk at Ser345 (9), Chk2 at Thr68 (10) and p53 (11). The Chk kinases inactivate cdc25 via phosphorylation at Ser216, blocking the activation of cdc2.

1. Nurse, P. (1997) Cell 91, 865-7.
2. Norbury, C. and Nurse, P. (1992) Annu Rev Biochem 61, 441-70.
3. Watanabe, N. et al. (1995) EMBO J 14, 1878-91.
4. Sherr, C.J. (1996) Science 274, 1672-7.
5. Dyson, N. (1998) Genes Dev 12, 2245-62.
6. Kitagawa, M. et al. (1996) EMBO J 15, 7060-9.
7. Lundberg, A.S. and Weinberg, R.A. (1998) Mol Cell Biol 18, 753-61.
8. Harbour, J.W. et al. (1999) Cell 98, 859-69.
9. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39.
10. Matsuoka, S. et al. (2000) Proc Natl Acad Sci USA 97, 10389-94.
11. Tibbetts, R.S. et al. (1999) Genes Dev 13, 152-7.

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

2348:

Flow, Immunofluorescence, Western Blotting

2661:

Flow, Immunofluorescence, Immunoprecipitation, Western Blotting

8516:

IHC / Paraffin, Immunofluorescence, Immunoprecipitation, Western Blotting

9111:

ELISA Peptide, Immunoprecipitation, Western Blotting

9284:

ChIP Agarose, ChIP Magnetic, Immunofluorescence, Immunoprecipitation, Western Blotting

9301:

Immunoprecipitation, Western Blotting

Companion Products

• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7076 Anti-mouse IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9786 Mismatch Repair Antibody Sampler Kit
• 9919 Phospho-p53 Antibody Sampler Kit
• 9931 Phospho-Chk1/2 Antibody Sampler Kit
• 8344 MRN Complex Antibody Sampler Kit
• 9947 DNA Damage Antibody Sampler Kit
• 8342 UV Induced DNA Damage Response Antibody Sampler Kit

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