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/ ) oncogene” Oh cc? RESEARCH PRODUCTS c-H-ras (Ab-1) Cat# OP23, OP23L Background: The human ras gene family consists of three identified members, H, K and N-ras, encoding proteins of 188-189 amino acids and 21,000 (p21) molecular weight (1,2). Human H- and K-ras are the homologues of v-H- and v-K-ras sequences originally isolated from Harvey and Kirsten strains of rat sarcoma viruses (3.4). Normal human cellular ras genes can be activated to oncogenes by mutations occurring in codons 12, 13 and 61; such mutated, activated and transforming ras genes have been identified and isolated from human tumors and cultured tumor cells (for review see 5). Although the expression patterns of ras proto-oncogene proteins in normal human tissues are known (6), similar information for activated ras oncogene encoded p21's and their relevance to human disease diagnosis and prognosis is still emerging (7, 8, 9). Origin: Clone F235-1.7.1 is a mouse monoclonal antibody generated by immunizing BALB/c mice with recombinant p21 protein and fusing with P3X63 Ag8.653 myeloma cells. Characteristics: Isotype: IgGi« Epitope: within residues 54-188 Species human] mouse rat other Reactivity legend: Y=yes NT=not tested Applications: Immuno- amount label positive Precipitation* control Frozen amount positive negative Sections control control pe (Ab-1) Paraffin amount detergent enzyme positive negative Sections control control trp (Ab-1) Western amount chemi. colori. positive Blotting* luminescent metric control 10 pg/ml. Immuno- amount positive fluorescence control legend: Y=yes NT=not tested *See Comments Oncogene Research Products Cat# OP23, OP23L Rev. 22-Sep-98 BOF G \\ eat

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How Supplied: 100 pg or 200 pg (Cat# OP23) of purified antibody in 1.0 mL of 0.05 M sodium phosphate buffer containing 0.1% sodium azide and 0.2% gelatin; or 100 pg (Cat# OP23L) purified antibody lyophilized from a volatile buffer with 100 pg of BSA. We recommend resuspending the lyophilized antibody with sterile phosphate buffered saline (PBS), pH 7.4, or sterile 20 mM Tris-saline (20 mM Tris containing 0.15 M NaCl), pH 7.4, to yield a final concentration of 100 pg/mL; product will be more stable if 0.1% sodium azide is included (do not add azide if antibody is to be used with viable cells). Lyophilized antibody should be resuspended at 4°C with occasional gentle mixing for at least two hours. Storage: Store Cat# OP23 (in solution) at 4°C; do not freeze. Store Cat# OP23L (lyophilized) at 4°C until reconstituted, then store in aliquots at -20°C or at 4°C with 0.1% azide; freezing of aliquots is best for storage of reconstituted product for longer than a month, but repetitive freezing and thawing should be avoided. If stored under proper conditions, product guaranteed until expiration date stated. Comments: For immunoprecipitation, use 5 pg Cat# OP23 per sample with 45 pL protein G plus agarose. The level of expression of p21'45 is variable in different tissues. For this reason, we recommend a concentration step prior to western blot analysis to obtain optimal results. A doublet may be seen due to farnysylation. c-H-ras (Ab-1) will react to C-H-ras and, weakly, to v-H-ras, it does not detect either c-K-ras or c-N-ras p21s under the conditions tested. Purified p21 ras proteins are also available for western blotting standards. Suggested starting concentrations are provided. Antibodies should be titrated for optimal results in individual systems. References: 2. Taparowsky, E., et al., Cell 34: 581-586, 1983. 6. Furth, M. E., etal, Oncogene 1: 47-58, 1987. 8. Sidransky, D., et al., Science 256: 102-105, 1992. © 1996 Calbiochem-Novabiochem, International Cat## OP23, OP23L Rev. 22-Sep-98 BOF ONCOGENE RESEARCH PRODUCTS ORDERING INFORMATION

84 Rogers Street, Cambridge, MA 02142 Research reagents may be ordered by mail at

Tel: 617 577-1639 800 662-2616 the address shown at left, by telephone, Fax: 617 577-8015 800 828-4871 E-mail, or fax or from your local E-Mail: customer.service@oncresprod.com Oncogene Research Products supplier. For research use only, not for use in diagnostic procedures V, ¥ — IAEA AtKAteA