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
a synthetic peptide (TAENPEYLGLDVPV) corresponding to amino acids 1242-1255 from the C-terminal domain of human c-ErbB2/c-Neu
Human
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.
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.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Standard Handling (A)
| biological source | mouse |
| Quality Segment | 100 |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 3B5, monoclonal |
| form | lyophilized |
| does not contain | preservative |
| species reactivity | mouse, human |
| manufacturer/tradename | Calbiochem® |
| storage condition | OK to freeze |
| dilution | (Frozen Sections (2.5 µg/mL) Immunoblotting (2.5 µg/mL, Immunofluorescence (2.5 µg/mL) Immunoprecipitation (1 µg per sample, Paraffin Sections (2.5 µg/mL, pressure cooker pre-treatment required, ) |
| isotype | IgG1 |
| shipped in | ambient |
| storage temp. | 2-8°C |
| target post-translational modification | unmodified |
| Gene Information | human ... ERBB2(2064) |
