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General description
Anti-EGFR (Ab-1), mouse monoclonal, clone 528, recognizes EGF receptor protein in A431 cells and normal skin. It is validated for use in ELISA, IF, IP, IHC (frozen,paraffin) & neutralization studies.
Purified mouse monoclonal antibody generated by immunizing Balb/c mice with the specified immunogen and fusing splenocytes with NS-1 mouse myeloma cells (see application references). Recognizes the ~170 kDa EGFR protein.
Recognizes the EGF receptor protein in A431 cells and normal skin.
Immunogen
partially purified, human EGFR from human A431 cells
Application
Flow Cytometry (5 µg/ml)
Frozen Sections (2.5 µg/ml)
Immunofluorescence (5 µg/ml)
Immunoprecipitation (1 µg/sample)
Neutralization Studies (see comments and application references)
Paraffin Sections (see application references)
Physical form
Lyophilized from 20 mM ammonium bicarbonate solution, 0.1 mg FAF-BSA.
Preparation Note
We recommend resuspending the lyophilized antibody with sterile phosphate buffered saline (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; product will be more stable if 0.1% sodium azide is included (do not add azide if antibody is to be used with viable cells). Lyophilized antibody should be resuspended at 4°C with occasional gentle mixing for at least 2 h. Following reconstitution, refrigerate (4°C) for short-term storage or aliquot and freeze (-20°C) for long-term storage.
Analysis Note
Positive Control
A-431 cells or skin tissue
Other Notes
Hunter T. 1984. Nature311, 414.
Masui, H., et al. 1984. Cancer Res.44, 1002.
Reynolds, F. H., Jr., et al. 1981. Nature 292, 259.
Ushiro, H. and Cohen, S. 1980. J. Biol. Chem.255, 8363.
This antibody binds to A431, HeLa, and other human cells but not to rodent cells. This antibody inhibits EGF binding to its receptor and is an antagonist of in vivo EGF-stimulated tyrosine kinase activity. Also inhibits A431 tumor formation in nude mice. For neutralization studies, use Cat. No. GR01L; Cat. No. GR01 contains azide, which is toxic to cells. Antibody should be titrated for optimal results in individual systems.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Standard Handling (A)
| biological source | mouse |
| Quality Level | 100 |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 528, monoclonal |
| form | lyophilized |
| does not contain | preservative |
| species reactivity | human |
| should not react with | rat, mouse |
| manufacturer/tradename | Calbiochem® |
| storage condition | OK to freeze |
| dilution | (Flow Cytometry (5 µg/mL) Frozen Sections (2.5 µg/mL) Immunofluorescence (5 µg/mL) Immunoprecipitation (1 µg/sample) Neutralization Studies Paraffin Sections ) |
| isotype | IgG2a |
| shipped in | ambient |
| storage temp. | 2-8°C |
| target post-translational modification | unmodified |
| Gene Information | human ... EGFR(1956) mouse ... Egfr(13649) rat ... Egf(25313) |
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