EGFR pTyr869 (incl. pos. control) Mouse Monoclonal Antibody [Clone ID: 12A3]

Specifications

Product Data
Clone Name	12A3
Applications	ELISA, IF, IHC, IP, WB
Recommend Dilution	ELISA: Use at 0.1 µg/ml (direct ELISA). Immunocytochemistry: Use at 1-10 µg/ml. Immunohistochemistry on Frozen and Paraffin Embedded Material. Immunoprecipitation: Use at 1-10 µg per 10e6 vanadate-treated A431 cells. Western Blot: 1 µg/ml for HRPO/ECL detection. Recommended buffer: Casein/Tween 20 based blocking and blot incubation buffer (cat# AS00002BU-N). Included Positive Control: Cell lysate delivered with this product (See Protocol below!).
Reactivity	Human, Mouse
Host	Mouse
Clonality	Monoclonal
Immunogen	Phosphopeptide conjugated to KLH Epitope: Phosphotyrosine 869
Specificity	Clone 12A3 specifically recognizes EGFR phosphorylated at Tyrosine 869 and detects EGFR activation after interaction with src kinases. Antibody AM00031PU-N does not crossreact with the highly homologues pTyr 877 of activated erbB2.
Formulation	PBS containing 0.09% Sodium Azide / PEG and Sucrose. State: Purified State: Lyophilized purified Ig fraction.
Reconstitution Method	Restore in 1 ml distilled water for 15 min at RT.
Purification	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 2-8°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	Mol. weight of antigen: 180 kD. A positive Control is provided (for details see protocol below). Protocol: Positive control cell lysate provided: HepG2 EGF Treated - recommended for immunoblotting Formulation: Lyophilized cell lysate from HepG2 cells. Serum starved cells were treated for 15 min. with EGF. 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. Applications: 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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