Analysis Note

Positive ControlSW480 or Y1 cells or normal skin tissue

Application

Frozen Sections (5 µg/ml)r>Immunoblotting (2.5 µg/ml, colormetric or chemiluminescence)r>Immunofluorescence (2.5 µg/ml)r>Immunoprecipitation (use Cat. No. OP01A or 1 µg antibody per sample)r>Paraffin Sections (5 µg/ml, saponin or pepsin pre-treatment required)r>Neutralization Studies (see application references)

General description

This Anti-v-H-Ras (Ab-1) Rat mAb (Y13-259) is validated for use in Frozen Sections, Immunoblotting, IF, IP, Paraffin Sections, Neutralization Studies for the detection of v-H-Ras (Ab-1).

Recognizes both phosphorylated and nonphosphorylated forms of v-H-Ras, and v-K-Ras in SW480 and Y1 cells and normal skin tissue. Also recognizes H, K, and N-Ras human translated products.

Purified rat monoclonal antibody generated by immunizing Sprague Dawlay rats with the specified immunogen and fusing splenocytes with Y3Ag1.2.3 rat myeloma cells. Recognizes the ~21 kDa Ras protein (see comments).

Immunogen

recombinant p21 Ras

Epitope: within amino acids 62-76

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Broadly reactive with the phosphorylated and non-phosphorylated forms of v-H-Ras and v-K-Ras (see application references) and recognizes the p21 translational products of the H-, K-, and N-ras human genes (see application references). This antibody does not cross-react with rap. Neutralizes the biological and biochemical activities of H-, K-, and N-ras p21 of vertebrates by binding to residues Glu-63, Ser-65, Ala-66, Met-67, Gln-70, and Arg-73 (see application references). 15 µl v-H-ras (Ab-1) agarose conjugate (Cat. No. OP01A) is recommended to be used as an immunoconcentrator prior to immunoblot analysis of p21^ras with another antibody. To immunoprecipitate with the unconjugated antibody, use Protein A with a rabbit anti-rat IgG or Protein G PLUS with a goat anti-rat IgG. Protein A alone will not immunoprecipitate rat IgG. A doublet may be detected corresponding to ras protein with and without farnesyl group. Antibody should be titrated for optimal results in individual systems.

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.

Packaging

100 µg in Plastic ampoule

Please refer to vial label for lot-specific concentration.

Physical form

In 0.05 M sodium phosphate buffer, 0.2% gelatin.

Warning

Toxicity: Standard Handling (A)