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|Antigen||Epidermal Growth Factor Receptor (EGFR) Antibodies|
|Reactivity||Human, Mouse (Murine) Alternatives|
|Conjugate||This EGFR antibody is un-conjugated Alternatives|
Enzyme Immunoassay (EIA), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Western Blotting (WB)
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Product Details anti-EGFR AntibodyTarget Details EGFR Application Details Handling Images
|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.|
|Cross-Reactivity (Details)||Species reactivity (tested):Human, Mouse.|
|Characteristics||Synonyms: Epidermal growth factor receptor, EGF Receptor, erbB-1, c-ErbB-1, Receptor tyrosine-proteinkinase erbB-1, Proto-oncogene c-ErbB-1|
|Purification||Subsequent Thiophilic Adsorption and Size Exclusion Chromatography.|
|Components||incl. positive Control|
|Immunogen||Phosphopeptide conjugated to KLHEpitope: Phosphotyrosine 869AA Sequence: E K E pY H A E|
|Plasmids, Primers & others|
Target Details EGFRProduct Details anti-EGFR Antibody Application Details Handling Images back to top
|Alternative Name||EGFR / ERBB1 (EGFR Antibody Abstract)|
|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: EGF Receptor, Epidermal growth factor receptor, Proto-oncogene c-ErbB-1, Receptor tyrosine-protein kinase erbB-1, c-ErbB-1, erbB-1|
|Pathways||NF-kappaB Signaling, RTK Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Stem Cell Maintenance, Hepatitis C, Positive Regulation of Response to DNA Damage Stimulus, Interaction of EGFR with phospholipase C-gamma, Thromboxane A2 Receptor Signaling, EGFR Downregulation|
Application DetailsProduct Details anti-EGFR Antibody Target Details EGFR Handling Images back to top
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 10^6 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! ).
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.
|Protocol||Positive control cell lysate provided: HepG2 EGF Treated - recommended for immunoblottingFormulation: Lyophilized cell lysate from HepG2 cells. Serum starved cells were treated for 15 min. withEGF. Restore by addition of 200 µl H2O. After complete solubilization add 200 µl 2x SDS-PAGEsample buffer, mix and incubate at 90°C for 5 min. Store in aliquots at -20°C. Avoidrepeated freezing and thawing. Applications: 20 µl of positive cell lysate correspond to ca. 80. 000 cells. Use 20 µl / lane (mini gel) forHRPO/ECL detection of the target proteins.|
|Restrictions||For Research Use only|
HandlingProduct Details anti-EGFR Antibody Target Details EGFR Application Details Images back to top
|Reconstitution||Restore in 1 mL distilled water for 15 min at RT.|
|Buffer||PBS containing 0.09 % Sodium Azide / PEG and Sucrose.|
|Precaution of Use||This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.|
|Storage Comment||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 3months. Avoid repeated freeze / thaw cycles!|
|Expiry Date||12 months|
ImagesProduct Details anti-EGFR Antibody Target Details EGFR Application Details Handling back to top