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 with Sp2/0.Ag14 cells (see application references). Recognizes the ~185 kDa c-ErbB2/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 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)
Physical form
Lyophilized from 20 mM ammonium bicarbonate Buffer, 100 µg FAF-BSA.
Preparation Note
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.
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.
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Yamamoto, T., et al. 1986. Nature319, 230.
Blick, M., et al. 1984. Blood64, 1234.
Schwab, M., et al. 1984. Cold Spring Harbor Laboratory2, 215.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Standard Handling (A)
