Anti-c-ErbB2/c-Neu (Ab-3) Mouse mAb (3B5)

Analysis Note

Negative ControlHepG2 cells or normal skin

Positive ControlSK-BR-3 cells or breast carcinoma tissue

Application

Frozen Sections (2.5 g/ml)Immunoblotting (2.5 g/ml, see application references)Immunofluorescence (2.5 g/ml)Immunoprecipitation (1 g per sample, see application references)Paraffin Sections (2.5 g/ml, pressure cooker pre-treatment required, see application references)

General description

Recognizes the ~185 kDa c-ErbB2/c-Neu protein in HEK293 cells.

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 with Sp2/0.Ag14 cells (see application references). Recognizes the ~185 kDa c-ErbB2/c-Neu protein.

Immunogen

a synthetic peptide (TAENPEYLGLDVPV) corresponding to amino acids 1242-1255 from the C-terminal domain of human c-ErbB2/c-Neu

Human

Legal Information

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

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 Laboratory2, 215.

Packaging

100 µg in Glass bottle

Physical form

Lyophilized from 20 mM ammonium bicarbonate Buffer, 100 µg FAF-BSA.

Reconstitution

Reconstitue 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 reconstituted at 4°C with occasional gentle mixing for at least 2 h. Store 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.

Warning

Toxicity: Standard Handling (A)