Description General description Anti-c-ErbB2/c-Neu (Ab-3), mouse monoclonal, clone 3B5, recognizes the ~185 kDa c-ErbB2/c-Neu protein in HEK293 cells. It is validated for WB, IF, IP, and IHC on frozen and paraffin sections. Purified mouse monoclonal antibody generated by immunizing BALB/c mice with the specified immunogen and fusing splenocytes with Sp2/0.Ag14 cells (see application references). Recognizes the ~185 kDa c-Erb2/c-Neu protein. Recognizes the ~185 kDa c-ErbB2/c-Neu protein in HEK293 cells. Immunogen Human a synthetic peptide (TAENPEYLGLDVPV) corresponding to amino acids 1242-1255 from the C-terminal domain of human c-ErbB2/c-Neu Application Frozen Sections (2.5 µg/ml) Immunoblotting (2-3 µg/ml, see application references) Immunofluorescence (2.5 µg/ml) Immunoprecipitation (1 µg/sample, see application references) Paraffin Sections (2.5 µg/ml, pressure cooker, heat or trypsin pre-treatment required, see application references) Physical form In 0.05 M sodium phosphate buffer, 0.2% gelatin. Packaging 100 μg in Plastic ampoule Please refer to vial label for lot-specific concentration. Warning Toxicity: Standard Handling (A) Analysis Note Negative Control HepG2 cells or normal skin Positive Control SK-BR-3 cells or breast carcinoma tissue Other Notes For paraffin sections, we recommend pre-treating with a pressure cooker, but trypsin and heat will also work well. Including 0.05% saponin during staining of paraffin sections may help reduce background. In one study ductal carcinomas in situ displaying large-cell, comedo growth type, were stained while none of 16 ductal carcinomas in situ of small-cell, papillary, or cribriform growth type were stained by this antibody. Antibody should be titrated for optimal results in individual systems. Mamcs, J.R., et al. 1994. Ann. Surg.219, 332. Tetu, B., et al. 1994. Cancer73, 2359. DiFiore, P.P., et al. 1987. Science237, 178. Slamon, D.J., et al. 1987. Science235, 177. Varley, J.M., et al. 1987. Oncogene1, 423. Bargmann, C.I., et al. 1986. Nature319, 226. Yamamoto, T., et al. 1986. Nature319, 230. Blick, M., et al. 1984. Blood64, 1234. Schwab, M., et al. 1984. Cold Spring Harbor Laboratory 2, 215.