EGFR pTyr1173 (incl. pos. control) Mouse Monoclonal Antibody [Clone ID: 9H2]

Specifications

Product Data
Clone Name	9H2
Applications	ELISA, IF, IHC, IP, WB
Recommend Dilution	ELISA: Use at 0.05 µg/ml. Western Blot: 0.5 µg/ml for HRPO/ECL detection. Recommended buffer: Casein/Tween 20 based blocking and blot incubation buffer. Use cell lysate delivered with this product as Positive Control (See Protocol below). Immunoprecipitation: Use at 1-10 µg per 106 vanadate-treated A431 cells. Immunocytochemistry: Use at 1-10 µg/ml. Immunohistochemistry on Frozen and Paraffin Embedded Material.
Reactivity	Canine, Human, Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	Phosphopeptide conjugated to KLH Epitope: phospho-Tyr1173 N A E pY L R V
Specificity	Clone 9H2 specifically interacts with the 1170 - N A E pY L R V motif corresponding to the major autophosphorylation site of Human EGFR. Antibody AM00042PU-N does not crossreact with the highly homologous pTyr1248 of acticated ErbB2.
Formulation	PBS containing 0.09% Sodium Azide, PEG and Sucrose State: Purified State: Lyophilized purified Ig fraction
Reconstitution Method	Restore with 1.0 ml H2O for 15 min at RT
Purification	Purified from Serum-Free Cell Culture Supernatant by Subsequent Thiophilic Adsorption and Size Exclusion Chromatography
Conjugation	Unconjugated
Storage Condition	For long-term storage, freeze lyophilizate upon arrival (-20°C). Upon reconstitution, aliquote and freeze in liquid nitrogen, reconstituted antibody can be stored frozen at -80°C up to 1 year. Thaw aliquots at 37°C. Thawed aliquots may be stored at 4°C up to 3 months. Avoid repeated freeze / thaw cycles!
Gene Name	epidermal growth factor receptor
Database Link	Entrez Gene 1956 Human UniProt ID P00533
Background	EGFR/erbB receptors are activated upon binding of EGF and EGF-related growth factors such as TGF alpha, beta-cellulin, Hb-EGF, HRG, or NRG. Binding of these ligands leads to receptor homo- and heterodimerization followed by autophosphorylation and activation of downstream signal transduction pathways (MAPK, PI3K/PKB, and STAT). In addition, EGFR becomes fully activated after phosphorylation of Y869 by src family kinases. Phosphorylation of Y1069 leads to association with cbl and subsequent receptor degradation. Phosphorylation of S1071 by CamKinase II leads to attenuation of kinase activity; phosphorylation of T678 (by PKC) and T693 (by MAPK, p38) interferes with receptor endocytosis/recycling.
Synonyms	Epidermal growth factor receptor, EGF Receptor, erbB-1, c-ErbB-1
Note	A Positive Control cell lysate is provided with this antibody (for details see Protocol below). Protocol: Provided Positive Control cell lysate: HepG2 Pervanadate Treated -recommended for Immunoblotting Formulation: Lyophilized cell lysate from HepG2 cells. Serum starved cells were treated for 15 min. with pervanadate. Restore by addition of 200 µl H2O. After complete solubilization add 200 µl 2x SDS-PAGE sample buffer, mix and incubate at 90°C for 5 min. Store in aliquots at -20°C. Avoid repeated freezing and thawing. Application: 20 µl of positive cell lysate correspond to ca. 80.000 cells. Use 20 µl / lane (mini gel) for HRPO/ECL detection of the target proteins. Please note: The lyophilized cell lysate contains SDS and is not recommended for applications with native proteins such as Immunoprecipitation !
Reference Data

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