Epidermal Growth Factor Receptor (erbB, EGFR), Neutralizing

Applications: \nSuitable for use in Western Blot, Immunoprecipitation, Immunohistochemistry, Immunocytochemistry and Neutralization. Other applications have not been tested.\n\nRecommended Dilutions:\nImmunohistochemistry: Acetone-fixed tissues. \nNeutralization of Biological Activity: 10ug/ml causes a 74% reduction in cell number in a 5 day cell proliferation assay using A431 cells. \nWestern Blot: 0.5-2ug/ml detects non-reduced EGF receptor in RIPA lysates from non-stimulated human A431 cells. Non-stimulated A431 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with E3375-14 (1ug/ml). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemilum-inescence detection system.\nImmunoprecipitation: 4ug immunoprecipitates the EGF receptor from 500ug of non-stimulated A431 RIPA lysate. \nImmunocytochemistry: 10ug/ml shows positive immunostaining for the EGF receptor in A431 cells fixed with 4% para-formaldehyde.\nOptimal dilutions to be determined by the researcher.\n\nBiological Activity: \nInhibits A431 cell growth. Recognizes an antigenic determinant located on the extracellular domain of the receptor.\n\nPositive Antigen Control: \nA0001-50: A431 Cells, Non-Stimulated Use 20ug per lane for minigels.\n\nStorage and Stability:\nLyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute with sterile ddH2O.. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

E3375-14

Size

250ug

Applications

ICC, IHC, IP, WB

Hosts

Mouse

Reactivities

Hum

Form

Supplied as a lyophilized powder from 0.1M Tris-glycine, pH 7.4, 0.15M sodium chloride. Reconstitute with 250ul sterile ddH2O. Further dilute with PBS, 5mg/ml BSA.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG1

References

Shaw, et al., J. Biol. Chem. 266: 21114, 1991. Kawamoto,et al., In Vitro C. Dev. Biol. 28A: 782, 1992. Johnson, et al., J. Biol. Chem. 268: 2924, 1993. Lillien, et al., Nature 377: 158, 1995.

Additional Info

Recognizes the external domain of EGF receptor on all human cells at ~180kD. Competes with EGF and TGF-a for binding on human cells. Recognizes both phosphorylated and non-phosphorylated human EGF receptors. Species crossreactivity: does not react with EGF receptors on mouse or rat cells.