Product Pathways - Cell Cycle / Checkpoint
p21 Waf1/Cip1 (DCS60) Mouse mAb #2946
PhosphoSitePlus® protein, site, and accession data: p21Cip1
| 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
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.
Western Blotting
Western blot analysis of extracts from 293, MCF-7 and COS cells, using p21 Waf1/Cip1 (DCS60) Mouse mAb.
Western Blotting
Western blot analysis of extracts from 293 and Hela cells, transfected with control (-) or p21 Waf1/Cip1 (+) siRNA. p21 Waf1/Cip1 was detected using p21 Waf1/Cip1 (DCS60) Mouse Monoclonal Antibody #2946, p42 MAPK was detected using p42 MAPK (3A7) Mouse Monoclonal Antibody #9107. The p21 Waf1/Cip1 monoclonal antibody confirms silencing of p21 expression, and the p42 monoclonal antibody is used to control for loading and specificity of p21 Waf1/Cip1 siRNA.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear and cytoplasmic localization, using p21 Waf1/Cip1 (DCS60) Mouse mAb.
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).
- Pestell, R.G. et al. (1999) Endocrine Rev. 20, 501-534.
- Cheng, J. et al. (1999) EMBO J. 18, 1571-1583.
- Flores-Rozas, H. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 8655-8659.
- Wang, Y. and Prives, C. (1995) Nature 376, 88-91.
- 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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For Research Use Only. Not For Use In Diagnostic Procedures.