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p21 Waf1/Cip1 (DCS60) Mouse mAb #2946

Applications	Reactivity	Sensitivity	MW (kDa)	Isotype
W IHC-P	H	Endogenous	21	Mouse IgG2a

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

2946:

Immunoprecipitation, Western Blotting

Specificity / Sensitivity

p21 Waf1/Cip1 (DCS60) Mouse mAb detects endogenous levels of total p21 protein. The antibody does not cross-react with other cdk inhibitors.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant human p21 corresponding to the amino-terminal portion of p21.

Background

The tumor suppressor protein p21 Waf1/Cip1 acts as an inhibitor of cell cycle progression. It functions in stoichiometric relationships forming heterotrimeric complexes with cyclins and cyclin-dependent kinases. In association with CDK2 complexes, it serves to inhibit kinase activity and block progression through G1/S (1). However, p21 may also enhance assembly and activity in complexes of CDK4 or CDK6 and cyclin D (2). The carboxy-terminal region of p21 is sufficient to bind and inhibit PCNA, a subunit of DNA polymerase, and may coordinate DNA replication with cell cycle progression (3). Upon UV damage or during cell cycle stages when cdc2/cyclin B or CDK2/cyclin A are active, p53 is phosphorylated and upregulates p21 transcription via a p53-responsive element (4). Protein levels of p21 are downregulated through ubiquitination and proteasomal degradation (5).

1. Pestell, R.G. et al. (1999) Endocrine Rev. 20, 501-534.
2. Cheng, J. et al. (1999) EMBO J. 18, 1571-1583.
3. Flores-Rozas, H. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 8655-8659.
4. Wang, Y. and Prives, C. (1995) Nature 376, 88-91.
5. Sheaff, R.J. et al. (2000) Cell 5, 403-410.

Application References

• Thullberg, M. et al. (2000) Monoclonal antibody probes for p21 Waf1/Cip1 and the INK4 family of cyclin-dependent kinase inhibitors. Hybridoma 19 (1), 63-72. Applications: IC-IF Western Blotting
• Zhu, H. et al. (2004) Induction of S-phase arrest and p21 overexpression by a small molecule 2[[3-(2,3-dichlorophenoxy)propyl] amino]ethanol in correlation with activation of ERK. Oncogene 23, 4984-4992. Applications: Western Blotting
• Chang, S.F. et al. (2008) Proc Natl Acad Sci U S A 105, 3927-32. Applications: Western Blotting
• Zhang, S. et al. (2009) Mol Cancer Res 7, 570-80. Applications: IC-IF Western Blotting
• Hawkes, W.C. et al. (2012) J Cell Biochem 113, 61-9. Applications: Western Blotting
• Taft, R.J. et al. (2011) Epigenetics Chromatin 4, 13. Applications: Western Blotting
• Negrotto, S. et al. (2011) Cancer Res 71, 1431-41. Applications: Western Blotting

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Companion Products

• 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
• 8112 SignalStain^® Antibody Diluent
• 8125 SignalStain^® Boost IHC Detection Reagent (HRP, Mouse)
• 5425 Normal Goat Serum
• 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
• 8059 SignalStain^® DAB Substrate Kit

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