New p21 Monoclonal Antibodies

Our new p21 monoclonal antibodies, clones SX118 and SXM30, are more versatile in terms of species recognition and applications than those previously introduced.

Clone SX118 reacts with human, rat and mouse p21.(1) The epitope has been mapped to the last 20 amino acids (inclusive of residues 145-164) of human p21, one of the most conserved regions between human and mouse p21. Reaction of the antibody with overlapping peptides fragments suggest that the epitope may be further mapped to residues 145-156 (TSMTDFYHSKRR). This sequence overlaps with the sequence of p21 (KRRQTSMTDFYH) which is responsible for the specific interaction of p21 with PCNA. Purified GST-human p21 recombinant fusion protein was used as immunogen. The antibody was purified from ascites or hybridoma tissue culture supernatant by Protein G affinity chromatography.

Clone SXM30 reacts with human, rat and mouse p21. With regards to cross-species recognition, SXM30 has similar properties to SX118.(1) Purified GST-human p21 recombinant fusion protein was used as immunogen. The antibody was purified from ascites or hybridoma tissue culture supernatant by Protein G affinity chromatography.

APPLICATIONS

The antibodies are useful for immunoprecipitation (1-2 µg/one million cells),(1,2) western blot analysis (1-2 µg/ ml),(1,2) flow cytometric analysis (titrate between 0.06-1 µg/1 x 10 million cells), indirect immunofluorescence microscopy of tissue-cultured cells,(1) immunohistochemistry of frozen tissue sections (1­5 µg/ml), and immunohistochemistry of paraffin-embedded tissue sections (20 µg/ml).(1) MCF-7 human breast carcinoma cells (ATCC HTB 22), HL-60 human leukemia cells (ATCC CCL 240), and EL4 mouse lymphoma cells (ATCC TIB 39) are suggested as positive controls. In immunohistochemistry, staining is primarily nuclear.(1) Tonsil and colon are suggested as positive controls for tissue staining. Refer to Fredersdorf et al., for information regarding induction of p21 expression, and for p21 distribution in different types of human tissues.(1) Store the antibodies at 4°C.

Cat. No. Description Clone Isotype Size
65951A Purified anti-p21 SX118 Mouse IgG1 0.1mg
65961A Purified anti-p21 SXM30 Mouse IgG1 0.1mg

For Research Use Only. Not For Diagnostic or Therapeutic Use.

REFERENCES

  1. Fredersdorf, S., A.W. Milne, P.A. Hall, and Z. Lu. 1996. Characterization of a panel of novel anti-p21Waf1/Cip1 expression in normal human tissues. Am. J. Pathol. 148:825-835.
  2. Mazars, R.G., and S.J. Parmjit. 1997. Expression of p24, a novel p21Waf1/Cip1/Sdi1 -related protein, correlates with measurement of the finite proliferative potential of rodent embryo fibroblasts. Proc. Natl. Acad. Sci. USA 94:151-156.
  3. Huppi, K., D. Siwarski, J. Doskik, P. Michieli, S. Reed, B. Mock, D. Givol, and J.F. Mushinki. 1994. Molecular cloning, sequencing, chromosomal localization and expression of mouse p21 (Waf1). Oncogene 9:3017-3020.

PharMingen supplies a wide range of antibodies for cell cycle and tumor suppressor protein research, including:

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