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324865 Sigma-AldrichPhosphoDetect™ Anti-EGFR (pTyr⁸⁴⁵) Rabbit pAb

This PhosphoDetect™ Anti-EGFR (pTyr⁸⁴⁵) Rabbit pAb is validated for use in Immunoblotting for the detection of EGFR (pTyr⁸⁴⁵).

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

SDS
CoA
References
Data Sheet

Synonyms: Anti-Epidermal Growth Factor Receptor (pTyr⁸⁴⁵)

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Overview

Replacement Information

Key Spec Table

Species Reactivity	Host	Antibody Type
H	Rb	Polyclonal Antibody

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Description
Overview	Recognizes the ~170 kDa EGFR protein phosphorylated at Tyr⁸⁴⁵ in EGF-treated A431 cells. Note: 1 T = 1 test. Sufficient for 10 Western miniblots.
Catalogue Number	324865
Brand Family	Calbiochem®
Synonyms	Anti-Epidermal Growth Factor Receptor (pTyr⁸⁴⁵)
Application Data	Detection of human epidermal growth factor receptor, phosphorylated on Tyr⁸⁴⁵, by immunoblotting. Samples: Whole cell lysate from A431 cells left unstimulated (lane 1) or stimulated with EGF (lanes 2-5). Primary antibody: PhosphoDetect™ Anti-Epidermal Growth Factor Receptor (pTyr⁸⁴⁵) Rabbit pAb (Cat. No. 324865) (1:1000) pre-incubated without peptide (lanes 1, 2), with non-phosphopeptide corresponding to the immunogen (lane 3), a generic phosphotyrosine-containing peptide (lane 4), or phosphopeptide immunogen (lane 5). Detection: chemiluminescence.

References
References	Sieg, D.J. et al. 2000. Nat. Cell. Biol. 2, 249. Barbier, A.J. et al. 1999. J. Biol. Chem. 274, 14067. Biscardi, J.S., et al. 1999. J. Biol. Chem. 274, 8335. Hashimoto, A. et al. 1999. J. Biol. Chem. 274, 20139 Poppleton, H.M. et al. 1999. Arch. Biochem. Biophys. 363, 227.

Product Information
Form	Liquid
Formulation	In Dulbecco's PBS (w/o Mg²⁺ and Ca²⁺), 1 mg/ml BSA, 50% glycerol, pH 7.4.
Positive control	A431 cells stimulated with EGF
Preservative	≤0.1% sodium azide
Quality Level	MQ100

Applications
Key Applications	Immunoblotting (Western Blotting)
Application Notes	Immunoblotting (1:1000)
Application Comments	The immunogen sequence is 100% conserved in mouse and rat and 93% conserved in chicken EGFR, but cross-reactivity has not been tested. When diluted to 1:1000 there is sufficient antibody to perform 10 miniblots (10 T) at 10 ml per blot. Variables associated with assay conditions will dictate the proper working dilution.

Biological Information
Immunogen	a synthetic phosphopeptide corresponding to amino acids surrounding the Tyr⁸⁴⁵ phosphorylation site of human EGFR
Host	Rabbit
Isotype	IgG
Species Reactivity	Human
Antibody Type	Polyclonal Antibody

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements

Storage and Shipping Information
Ship Code	Dry Ice Only
Toxicity	Standard Handling
Storage	≤ -70°C
Avoid freeze/thaw	Avoid freeze/thaw
Do not freeze	Ok to freeze
Special Instructions	Following initial thaw, aliquot and freeze (-70°C).

Packaging Information

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
324865-10T	04055977215816

Documentation

PhosphoDetect™ Anti-EGFR (pTyr⁸⁴⁵) Rabbit pAb SDS

Title
Safety Data Sheet (SDS)

PhosphoDetect™ Anti-EGFR (pTyr⁸⁴⁵) Rabbit pAb Certificates of Analysis

Title	Lot Number
324865	Enter Lot Number

References

Reference overview
Sieg, D.J. et al. 2000. Nat. Cell. Biol. 2, 249. Barbier, A.J. et al. 1999. J. Biol. Chem. 274, 14067. Biscardi, J.S., et al. 1999. J. Biol. Chem. 274, 8335. Hashimoto, A. et al. 1999. J. Biol. Chem. 274, 20139 Poppleton, H.M. et al. 1999. Arch. Biochem. Biophys. 363, 227.

Data Sheet

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 (pTyr⁸⁴⁵)
Application	Immunoblotting (1:1000)
Application Data	Detection of human epidermal growth factor receptor, phosphorylated on Tyr⁸⁴⁵, by immunoblotting. Samples: Whole cell lysate from A431 cells left unstimulated (lane 1) or stimulated with EGF (lanes 2-5). Primary antibody: PhosphoDetect™ Anti-Epidermal Growth Factor Receptor (pTyr⁸⁴⁵) Rabbit pAb (Cat. No. 324865) (1:1000) pre-incubated without peptide (lanes 1, 2), with non-phosphopeptide corresponding to the immunogen (lane 3), a generic phosphotyrosine-containing peptide (lane 4), or phosphopeptide immunogen (lane 5). Detection: chemiluminescence.
Description	Rabbit polyclonal antibody adsorbed against non-phosphopeptide corresponding to the immunogen phosphorylation site and a generic tyrosine phosphorylated peptide, followed by immunoaffinity chromatography. Recognizes the ~185 KDa EGFR protein phosphorylated on Tyr⁸⁴⁵.
Background	Epidermal Growth Factor (EGF) is responsible for a variety of biological effects including mitogenesis and differentiation. The EGF receptor (EGFR) has three domains: the extracellular, transmembrane, and cytoplasmic domain. Upon the binding of EGF to the extracellular domain, dimerization of the receptor occurs and the cytoplasmic domain displays tyrosine kinase activity. This leads to the autophosphorylation of the ~170 kDa glycoprotein on various residues within the cytoplasmic domain as well as the phosphorylation of intracellular substrates. Tyr⁸⁴⁵ is an autophosphorylation site within EGFR. Phosphorylation of tyrosine 845 is mediated by non-receptor tyrosine kinase c-Src.
Host	Rabbit
Immunogen	a synthetic phosphopeptide corresponding to amino acids surrounding the Tyr⁸⁴⁵ phosphorylation site of human EGFR
Isotype	IgG
Species	human
Positive control	A431 cells stimulated with EGF
Form	Liquid
Formulation	In Dulbecco's PBS (w/o Mg²⁺ and Ca²⁺), 1 mg/ml BSA, 50% glycerol, pH 7.4.
Preservative	≤0.1% sodium azide
Comments	The immunogen sequence is 100% conserved in mouse and rat and 93% conserved in chicken EGFR, but cross-reactivity has not been tested. When diluted to 1:1000 there is sufficient antibody to perform 10 miniblots (10 T) at 10 ml per blot. Variables associated with assay conditions will dictate the proper working dilution.
Storage	Avoid freeze/thaw ≤ -70°C
Do Not Freeze	Ok to freeze
Special Instructions	Following initial thaw, aliquot and freeze (-70°C).
Toxicity	Standard Handling
References	Sieg, D.J. et al. 2000. Nat. Cell. Biol. 2, 249. Barbier, A.J. et al. 1999. J. Biol. Chem. 274, 14067. Biscardi, J.S., et al. 1999. J. Biol. Chem. 274, 8335. Hashimoto, A. et al. 1999. J. Biol. Chem. 274, 20139 Poppleton, H.M. et al. 1999. Arch. Biochem. Biophys. 363, 227.

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