		Technical Data
E3375-14	Epidermal Growth Factor Receptor (erbB, EGFR), Neutralizing

Description:
Applications:
Suitable for use in Western Blot, Immunoprecipitation, Immunohistochemistry, Immunocytochemistry and Neutralization. Other applications have not been tested.

Recommended Dilutions:
Immunohistochemistry: Acetone-fixed tissues.
Neutralization of Biological Activity: 10ug/ml causes a 74% reduction in cell number in a 5 day cell proliferation assay using A431 cells.
Western 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.
Immunoprecipitation: 4ug immunoprecipitates the EGF receptor from 500ug of non-stimulated A431 RIPA lysate.
Immunocytochemistry: 10ug/ml shows positive immunostaining for the EGF receptor in A431 cells fixed with 4% para-formaldehyde.
Optimal dilutions to be determined by the researcher.

Biological Activity:
Inhibits A431 cell growth. Recognizes an antigenic determinant located on the extracellular domain of the receptor.

Positive Antigen Control:
A0001-50: A431 Cells, Non-Stimulated Use 20ug per lane for minigels.

Storage and Stability:
Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile ddH2O.. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 12 months at -20°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.

Type	Isotype	Clone	Grade
Mab	IgG1	0.T.37	Affinity Purified

Size		Storage	Shipping	Source	Host
250ug		-20°C	Blue Ice	Human	Mouse

Concentration:

~1mg/ml

Immunogen:

Human A431 membrane proteins.

Purity:

Purified by Protein G affinity chromatography.

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.

Specificity:

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.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

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.