Analysis Note

Negative ControlHT1080 cells

Positive ControlHL-60 cells or lung carcinoma

Application

Flow Cytometry (20 µg/ml or use Cat. No. OP10F)r>Frozen Sections (2-10 µg/ml)r>Immunoblotting (1-5 µg/ml, see application references)r>Immunofluorescence (1-5 µg/ml or use Cat. No. OP10F)r>Immunoprecipitation (1 µg/sample)r>Chromatin Immunoprecipitation (5 µg/ml, see application references)r>Paraffin Sections (not recommended)

General description

Purified mouse monoclonal antibody generated by immunizing BALB/c mice with the specified immunogen and fusing splenocytes with SP2/0 mouse myeloma cells. Recognizes the ~64-67 kDa (apparent MW) c-Myc protein.

Recognizes a ~60-67 kDa c-Myc protein in HL60 cells and lung carcinoma tissue.

This Anti-c-Myc (Ab-1) Mouse mAb (9E10) is validated for use in FC, Frozen Sections, Immunoblotting, IF, IP, Chromatin IP, Paraffin Sections for the detection of c-Myc (Ab-1).

Immunogen

Epitope: within amino acids 410-419

a synthetic peptide (AEEQKLISEEDLLRKRREQLKHKLEQLRNSCA) corresponding to amino acids 408-439 of human c-Myc

Human

Legal Information

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

Other Notes

LeGouy, E., et al. 1987. In Nuclear Oncogenes, Cold Spring Harbor Laboratory, 144.Cole, M.D. 1986. Ann. Rev. Gen.20, 361.Nisen, P.D., et al. 1986. Cancer Res.46, 6217.Nau, M.M., et al. 1985. Nature318, 69.Persson, H., et al. 1984. Science225, 687.Alitalo, K., et al. 1983. Proc. Natl. Acad. Sci. USA80, 1707.

The c-Myc protein is extremely labile. c-Myc degradation can be as a result of freeze/thaw cycles, thus, we recommend using fresh lysates with a cocktail of protease inhibitors. c-Myc is often destroyed during the processing of paraffin sections and can therefore be difficult to detect in this application. This antibody recognizes the c-Myc protein and its cleavage products. The sequence of c-Myc predicts a ~45 kDa protein, but c-Myc migrates under reducing conditions as a ~64-67 kDa band. This antibody will not detect v-Myc but may react weakly to rodent c-Myc when used at high concentrations (10 µg/ml). An extra ~34-40 kDa band may be detected in a immunoblot. May also be used in immunofluorescence as well as the detection and purification of recombinant proteins tagged with the Myc epitope sequence EQKLISEEDL (see application references). The immunogen, c-Myc (Peptide-1) is also available for competition studies (Cat. No. PP06). Antibody should be titrated for optimal results in individual systems.

Packaging

200 µg in Plastic ampoule

Physical form

Lyophilized from a volatile buffer, 100 µg BSA.

Reconstitution

Reconstitute the lyophilized antibody with sterile 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 µg/ml. Lyophilized antibodies should be resuspended at 4°C with occasional gentle mixing for at least 2 h.

Warning

Toxicity: Standard Handling (A)