Description General description Purified mouse monoclonal antibody generated by immunizing BALB/c mice with the specified immunogen and fusing splenocytes with P3X63 Ag8.653 mouse myeloma cells. Recognizes the ~21 kDa c-H-Ras protein. Recognizes the ~21 kDa c-H-Ras protein in normal skin tissue and Rat1 cells. Does not react with c-K-Ras or c-N-Ras p21. A doublet may be seen due to farnesylation. This Anti-c-H-Ras (Ab-1) Mouse mAb (F235-1.7.1) is validated for use in Frozen Sections, Immunoblotting, IF, IP, Paraffin Sections for the detection of c-H-Ras (Ab-1). Immunogen Epitope: within amino acids 54-188 recombinant p21 Ras Application Frozen Sections (5 µg/ml) Immunoblotting (10 µg/ml) Immunofluorescence (2.5 µg/ml) Immunoprecipitation (5 µg/ml) Paraffin Sections (5 µg/ml, saponin and pepsin pre-treatment required) Physical form In 50 mM sodium phosphate buffer, 0.2% gelatin, pH 7.4. Packaging 100, 200 μg in Plastic ampoule Please refer to vial label for lot-specific concentration. Warning Toxicity: Standard Handling (A) Analysis Note Positive Control Normal skin tissue or ras 1 cells Other Notes For immunoprecipitation, use 5 µg of Cat. No. OP23 per sample with 45 µl protein G plus agarose. The level of expression of p21Ras is variable in different tissues. For this reason, we recommend a concentration step prior to immunoblotting analysis to obtain optimal results. A doublet may be seen due to farnysylation. Recognizes c-H-Ras and, weakly, v-H-Ras; does not recognize either c-K-Ras or c-N-Ras p21s under the conditions tested. Antibody should be titrated for optimal results in individual systems. Rodenhuis, S. et al. 1992. Cancer Res. (Suppl) 52, 2665s. Sidransky, D., et al. 1992. Science256, 102. Bos, J.L. 1989. Cancer Res.49, 4682. Furth, M. E., et al. 1987. Oncogene1, 47. Kraus, M.H., et al. 1984. Proc. Natl. Acad. Sci. USA81, 5384. Shimizu, K., et al. 1983. Proc. Natl. Acad. Sci. USA80, 2112. Taparowsky, E., et al. 1983. Cell34, 581. Ellis, R.W., et al. 1981. Nature292, 506. Shih, T.Y., et al. 1980. Nature287, 686.