OP24 ETC | Alldatasheet
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< ) oncogene” c-K-ras (Ab-1) Cat# OP24, OP24L 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 p2Is and their relevance to human disease diagnosis and prognosis is still emerging (7,8,9). K- ras mutations have been identified in several cancers to date including lung and colorectal tumors (10, IL). Origin: Clone 234-4.2 is a mouse monoclonal antibody generated by immunizing BALB/c mice with recombinant p21 protein and fusing with Ag8.653 myeloma cells. Characteristics: Isotype: IgG2ak Epitope: within amino acid residues 54-189 Reactivity legend: Y=yes N=no NT=not tested Applications: Sections control 10 ug/ml. precipitation control see 5s Met Y1 and comments i SW 480 cells Western amount chemi- colori- positive Blotting luminescent metric control comments SW 480 cells Sections 10 pgimt legend: Y=yes N=no Cat# OP24, OP24L Rev. 26-Feb-98 BOF Z a La IAEA AtKAteA
How Supplied: 100 ug or 200 yg (Cat# OP24) 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 yg (Cat# OP24L) 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 g/mL. Lyophilized antibodies should be resuspended at 4°C with occasional gentle mixing for at least two hours. Storage: Store Cat# OP24 (in solution) at 4°C; do not freeze. Store Cat# OP24L (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 long term storage of reconstituted product; repetitive freezing and thawing should be avoided. If stored under proper conditions, product guaranteed until expiration date stated. Comments: c-K-ras (Ab-1) reacts with c- and v-K-ras p21, but not with c-H-ras or c-N-ras p2ls. The level of expression of p21 ras is variable in different tissues. For this reason we recommend a concentration step prior to western blotting at 5 g/mL with 1% gelatin (see ref. 12 for complete protocol). For immunoprecipitation, use 5 1g OP24 per sample with 45 L protein A agarose. 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. 4. Shih, T-Y., etal, Nature 287: 686-691, 1980. 8. Sidransky, D., ef al., Science 256: 102-105, 1992. 11. Shaw, P., et a/., Oncogene 6: 2121-2128, 1991. © 1996 Calbiochem-Novabiochem, International Cat# OP24, OP24L Rev. 26-Feb-98 BOF ONCOGENE RESEARCH PRODUCTS ORDERING INFORMATION
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