324864 Sigma-AldrichPhosphoDetect™ Anti-EGFR (pTyr¹¹⁷³) Mouse mAb (9H2)

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision	01-August-2007 RFH
Synonyms	Anti-Epidermal Growth Factor Receptor (pTyr¹¹⁷³)
Application	ELISA (0.05 µg/ml) Immunoblotting (1 µg/ml, colorimetric; 0.1 µg/ml, chemiluminescence) Immunoprecipitation (1-10 µg/sample)
Application Data	Detection of human EGFR, phosphorylated on Tyr1173, by immunoblotting. Samples: Whole cell extracts of control (lane 1), EGF stimulated (lane 2) or pervanadate treated (lane 3) A549 cells. Primary antibody: PhosphoDetect™ Anti-EGFR (pTyr1173) Mouse mAb (9H2) (Cat. No. 324864) (0.5 µg/ml). Detection: chemiluminescence.
Description	Mouse monoclonal antibody purified from serum-free cell culture supernatant by thiophilic adsorption and size exclusion chromatography. Recognizes the ~170 kDa EGFR protein phosphorylated on Tyr1173. Supplied with a positive control lysate derived from 2 x 106 vanadate-treated HepG2 cells.
Background	Binding of EGF to the extracellular domain of the EGF-receptor results in receptor dimerization and autophosphorylation on tyrosine residues. The major site of tyrosine phosphorylation is tyrosine 1173.
Host	Mouse
Immunogen species	Human
Immunogen	a synthetic phosphopeptide (NAEpYLRV) corresponding to amino acids 1170-1176 of human EGFR
Clone	9H2
Isotype	IgG₁
Species	human, mouse, rat
Positive control	Vanadate-treated HepG2 cells (included)
Form	Lyophilized
Formulation	100 µg antibody lyophilized from 2X PBS, PEG, sucrose and 200 µl lyophilized control lysate.
Preservative	≤0.1% sodium azide (antibody only)
Comments	Does not cross-react with non-phosphorylated EGFR or with unrelated tyrosine-phosphorylated proteins. Use 0.5% BSA and 0.5% casein for blocking. For immunoblotting with the positive control lysate use 5 µl for chemiluminescence or 20 µl for colorimetric per mini-gel lane. Variables associated with assay conditions will dictate the proper working dilution.
Storage	-20°C
Do Not Freeze	Ok to freeze
Special Instructions	Reconstitute the antibody with 1 ml H₂O (room temperature, 15 min). Following reconstitution, refrigerate (4°C) for short-term storage or aliquot, flash-freeze in liquid nitrogen, and freeze (-70°C) for long-term storage. Stock solutions are stable for up to 3 months at 4°C or for up to 1 year at -70°C. Thaw aliquots at 37°C. Avoid freeze/thaw cycles of solutions. Reconstitute the positive control lysate in 200 µl H₂O. Add 200 µl SDS-PAGE sample buffer and incubate at 90°C for 5 min, aliquot, and freeze (-20°C).
Toxicity	Standard Handling