This Article Order Full text via Infotrieve Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Fredersdorf, S. Articles by Lu, X. Search for Related Content PubMed PubMed Citation Articles by Fredersdorf, S. Articles by Lu, X.

American Journal of Pathology, Vol 148, 825-835, Copyright © 1996 by American Society for Investigative Pathology

REGULAR ARTICLES

Characterization of a panel of novel anti-p21Waf1/Cip1 monoclonal antibodies and immunochemical analysis of p21Waf1/Cip1 expression in normal human tissues

S Fredersdorf, AW Milne, PA Hall and X Lu
Ludwig Institute for Cancer Research, St. Mary's Hospital Medical School, London, United Kingdom.

As a universal inhibitor of cyclin-dependent kinases and one of the target genes of the tumor suppresser p53, p21Waf1/Cip1 can act as a tumor suppresser through its ability to control cell cycle progression. To study the function of p21Waf1/Cip1 protein and to investigate its tissue distribution, a panel of anti-p21Waf1/Cip1 monoclonal antibodies was generated. These anti-p21Waf1/Cip1 monoclonal antibodies were initially raised against a GST-p21Waf1/Cip1 fusion protein produced in bacteria. Detailed characterization of the antibodies showed that they can specifically detect p21Waf1/Cip1 by immunoblotting, immunoprecipitation, and immunostaining. The specific induction of p21Waf1/Cip1 expression in response to gamma-radiation in cells containing p53 was also detected by these antibodies. The ability to detect p21Waf1/Cip1 expression in conventionally fixed tissue sections allowed us to investigate the distribution of p21Waf1/Cip1 in 23 different types of normal human tissues, and p21Waf1/Cip1 expression was found in most tissues. A close inverse relationship between p21Waf1/Cip1 expression and proliferation was seen in some tissues, including gastrointestinal tract. However, such association is not universal. In tissues such as lung, kidney, thyroid, pancreatic ducts and acini, and liver, despite the fact that most of the cells are quiescent, expression of p21Waf1/Cip1 was detected only in occasional epithelial cells. All these suggest that the expression of p21Waf1/Cip1 varies among different human tissues. Finally, epitope mapping of the anti-p21Waf1/Cip1 antibodies using a peptide library covering the entire p21Waf1/Cip1 protein sequence indicates that two of the antibodies recognize a region of p21Waf1/Cip1 close to that bound by proliferating cell nuclear antigen. These two monoclonal antibodies will therefore be additionally useful in further understanding the functions of p21Waf1/Cip1 both in vitro and in vivo.

This article has been cited by other articles:

				W. Zhang, D. Bergamaschi, B. Jin, and X. Lu Posttranslational modifications of p27kip1 determine its binding specificity to different cyclins and cyclin-dependent kinases in vivo Blood, May 1, 2005; 105(9): 3691 - 3698. [Abstract] [Full Text] [PDF]

				P. Muller, P. Ceskova, and B. Vojtesek Hsp90 Is Essential for Restoring Cellular Functions of Temperature-sensitive p53 Mutant Protein but Not for Stabilization and Activation of Wild-type p53: IMPLICATIONS FOR CANCER THERAPY J. Biol. Chem., February 25, 2005; 280(8): 6682 - 6691. [Abstract] [Full Text] [PDF]

				H. Wu, T. Hayashi, and M. Inoue Immunohistochemical Expression of p27 and p21 in Canine Cutaneous Mast Cell Tumors and Histiocytomas Vet. Pathol., May 1, 2004; 41(3): 296 - 299. [Abstract] [Full Text] [PDF]

				M. A. Sheard, S. Uldrijan, and B. Vojtesek Role of p53 in Regulating Constitutive and X-Radiation-Inducible CD95 Expression and Function in Carcinoma Cells Cancer Res., November 1, 2003; 63(21): 7176 - 7184. [Abstract] [Full Text] [PDF]

				G. P. Ilyin, D. Glaise, D. Gilot, G. Baffet, and C. Guguen-Guillouzo Regulation and role of p21 and p27 cyclin-dependent kinase inhibitors during hepatocyte differentiation and growth Am J Physiol Gastrointest Liver Physiol, June 9, 2003; 285(1): G115 - G127. [Abstract] [Full Text] [PDF]

				S. Hietanen, S. Lain, E. Krausz, C. Blattner, and D. P. Lane Activation of p53 in cervical carcinoma cells by small molecules PNAS, July 18, 2000; 97(15): 8501 - 8506. [Abstract] [Full Text] [PDF]

				Z Kolár, P G Murray, K Scott, A Harrison, B Vojtesek, and J Dusek Relation of Bcl-2 expression to androgen receptor, p21WAF1/CIP1, and cyclin D1 status in prostate cancer Mol. Pathol., February 1, 2000; 53(1): 15 - 18. [Abstract] [Full Text]

				C. Pfister, L. Moore, P. Allard, H. Larue, L. Lacombe, B. Tetu, F. Meyer, and Y. Fradet Predictive Value of Cell Cycle Markers p53, MDM2, p21, and Ki-67 in Superficial Bladder Tumor Recurrence Clin. Cancer Res., December 1, 1999; 5(12): 4079 - 4084. [Abstract] [Full Text] [PDF]

				T. Taniguchi, H. Endo, N. Chikatsu, K. Uchimaru, S. Asano, T. Fujita, T. Nakahata, and T. Motokura Expression of p21Cip1/Waf1/Sdi1 and p27Kip1 Cyclin-Dependent Kinase Inhibitors During Human Hematopoiesis Blood, June 15, 1999; 93(12): 4167 - 4178. [Abstract] [Full Text] [PDF]

				H. H. Cha, E. J. Cram, E. C. Wang, A. J. Huang, H. G. Kasler, and G. L. Firestone Glucocorticoids Stimulate p21 Gene Expression by Targeting Multiple Transcriptional Elements within a Steroid Responsive Region of the p21waf1/cip1 Promoter in Rat Hepatoma Cells J. Biol. Chem., January 23, 1998; 273(4): 1998 - 2007. [Abstract] [Full Text] [PDF]

				R. D. M. Steenbergen, J. N. Parker, S. Isern, P. J. F. Snijders, J. M. M. Walboomers, C. J. L. M. Meijer, T. R. Broker, and L. T. Chow Viral E6-E7 Transcription in the Basal Layer of Organotypic Cultures without Apparent p21cip1 Protein Precedes Immortalization of Human Papillomavirus Type 16- and 18-Transfected Human Keratinocytes J. Virol., January 1, 1998; 72(1): 749 - 757. [Abstract] [Full Text] [PDF]

				C. Ihling, G. Menzel, E. Wellens, J. S. Monting, H. E. Schaefer, and A. M. Zeiher Topographical Association Between the Cyclin-Dependent Kinases Inhibitor P21, p53 Accumulation, and Cellular Proliferation in Human Atherosclerotic Tissue Arterioscler. Thromb. Vasc. Biol., October 1, 1997; 17(10): 2218 - 2224. [Abstract] [Full Text]

				S. Fredersdorf, J. Burns, A. M. Milne, G. Packham, L. Fallis, C. E. Gillett, J. A. Royds, D. Peston, P. A. Hall, A. M. Hanby, et al. High level expression of p27kip1 and cyclin D1 in some human breast cancer cells: Inverse correlation between the expression of p27kip1 and degree of malignancy in human breast and colorectal cancers PNAS, June 10, 1997; 94(12): 6380 - 6385. [Abstract] [Full Text] [PDF]