Specification Please note DAKO Sheet Some information s Lot dependent Monoclonal mouse uman p Clone SX118 Code No M 7202 Lot 088 Edition 09.11.00 Introduction p21 is a protein encoded by the WAF1/Cip1 gene on chromosome 6p. p21 Ar p has been shown to inhibit the activity of several cyclin/cyclin-dependent kinase complexes and to block cell cycle progression (1). It was identified as a gene whose product was transcriptionally activated by wild-type but not mutant p53 protein, serving as a mediator of the cell cycle arrest function of p53( 2 ). In tumour cells that have lost the p53 protein or contain an altered form of p53, p21 levels are dramatically reduced or totally absent(2) This could lead to the passing of the G1 checkpoint, permitting the progression of the cell and oncogenesis. Because p21 appears to mediate several of the growth-reguatapl cycle in the presence of dNa alterations and in this way may result in genomic instability functions of p53, its expression would be predicted to reflect the functional status of p53 more precisely than p53 accumulation. The p21 AF l/cip protein can also be induced in a p53-independent manner and has been shown to play a role in differentiation (3) Furthermore, p21 r p can inhibit DNA replication by blocking the action of proliferating ell nuclear antigen(PCNA)(4). The expression of p21 has been studied immunohistochemistry in a wide range of human tumours including melanomas(5) pancreatic carcinomas(6), cervical carcinomas(7), thymomas(8), thyroid carcinomas( 9) breast carcinomas(10), head-and-neck carcinomas(11), colon carcinomas(12) and Hodgkins disease(13) Presentation Monoclonal mouse antibody supplied in liquid form as tissue culture supernatant(RPMI 1640 medium containing fetal calf serum) dialysed against 0.05 mol/L Tris/HCL, pH 7.2 containing 15 mmol/L NaN3 Total protein concentration: 11.1 g/L Storage 2-8°. Clone sx118.(14) Immunogen Purified glutathione S-transferase GST)-P27 WAF 1/Clp1 fusion protein Specificitylreactivity The antibody reacts with a 21 kDa protein in immunoblotting of MCF-7 cells and immunoprecipitates a 21 kDa band from lysates from SAOS-2 cells transfected with the full length p21 Ar cp protein(14). The antibody also reacts with p21 from mouse Application The antibody can be used in immunohistochemistry. Furthermore, the antibody produced by the clone SX118 has been shown to be applicable for immunoblotting an immunoprecipitation(14) Staining procedures Formalin-fixed and paraffin-embedded sections Can be used on formalin-fixed, paraffin-embedded tissue sections. Antigen retrieval, such as by heating in 10 mmol/L citrate buffer or in DAKO Target Retrieval Solution, code No. S1700, is mandatory. The slides should not be allowed to dry out during this treatment or during the following immunohistochemical staining procedureSpecification Sheet Please note Some information is Lot dependent. Monoclonal Mouse Anti-Human p21WAF1/Cip1 Clone SX118 Code No. M 7202 Lot 088. Edition 09.11.00 Introduction p21 is a protein encoded by the WAF1/Cip1 gene on chromosome 6p. p21WAF1/Cip1 has been shown to inhibit the activity of several cyclin/cyclin-dependent kinase complexes and to block cell cycle progression (1). It was identified as a gene whose product was transcriptionally activated by wild-type but not mutant p53 protein, serving as a mediator of the cell cycle arrest function of p53 (2). In tumour cells that have lost the p53 protein or contain an altered form of p53, p21 levels are dramatically reduced or totally absent (2). This could lead to the passing of the G1 checkpoint, permitting the progression of the cell cycle in the presence of DNA alterations and in this way may result in genomic instability and oncogenesis. Because p21 appears to mediate several of the growth-regulatory functions of p53, its expression would be predicted to reflect the functional status of p53 more precisely than p53 accumulation. The p21WAF1/Cip1 protein can also be induced in a p53-independent manner and has been shown to play a role in differentiation (3). Furthermore, p21WAF1/Cip1 can inhibit DNA replication by blocking the action of proliferating cell nuclear antigen (PCNA) (4). The expression of p21WAF1/Cip1 has been studied by immunohistochemistry in a wide range of human tumours including melanomas (5), pancreatic carcinomas (6), cervical carcinomas (7), thymomas (8), thyroid carcinomas (9), breast carcinomas (10), head-and-neck carcinomas (11), colon carcinomas (12) and Hodgkin’s disease (13). Presentation Monoclonal mouse antibody supplied in liquid form as tissue culture supernatant (RPMI 1640 medium containing fetal calf serum) dialysed against 0.05 mol/L Tris/HCl, pH 7.2 containing 15 mmol/L NaN3. Mouse Ig concentration: 390 mg/L. Isotype: IgG1, kappa. Total protein concentration: 11.1 g/L. Storage 2 - 8 °C. Clone SX118. (14). Immunogen Purified glutathione S-transferase (GST)-p21WAF1/Cip1 fusion protein. Specificity/reactivity The antibody reacts with a 21 kDa protein in immunoblotting of MCF-7 cells and immunoprecipitates a 21 kDa band from lysates from SAOS-2 cells transfected with the full length p21WAF1/Cip1 protein (14). The antibody also reacts with p21 from mouse. Application The antibody can be used in immunohistochemistry. Furthermore, the antibody produced by the clone SX118 has been shown to be applicable for immunoblotting and immunoprecipitation (14). Staining procedures Formalin-fixed and paraffin-embedded sections Can be used on formalin-fixed, paraffin-embedded tissue sections. Antigen retrieval, such as by heating in 10 mmol/L citrate buffer, pH 6.0, or in DAKO Target Retrieval Solution, code No. S 1700, is mandatory. The slides should not be allowed to dry out during this treatment or during the following immunohistochemical staining procedure. (2)