Peptide Competition. Extracts of A431 cells unstimulated (1) or stimulated with 200 ng/mL EGF for 15 minutes (2-5) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with the EGFR [pY1086] antibody for two hours at room temperature in a 1% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphotyrosine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab and quote;)2 anti-rabbit IgG HRP conjugate (Cat. no. ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to EGFR [pY1086] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show the induction of EGFR [pY1086] phosphorylation by the addition of EGF to this cell system.
|Tested species reactivity||Human|
|Published species reactivity||Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human EGFR that contains tyrosine 1086. The sequence is conserved in mouse and rat.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (IHC)||Assay Dependent|
|Western Blot (WB)||1:200-1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Epidermal Growth Factor Receptor (EGFR) is a 175 kDa transmembrane glycoprotein belonging to tyrosine kinase superfamily. It acts as a receptor for epidermal growth factor (EGF) family proteins. Binding of EGFR to its ligands causes autophosphorylation of tyrosine kinase followed by activation of signal transduction pathways connected to cell proliferation and differentiation. Tyrosine 1068 present within the cytoplasmic domain of the receptor is a major autophosphorylation site that allows binding of Grb2 and activation of the Ras-Raf-ERK1 and ERK2 signaling pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Hydrocortisone and indomethacin negatively modulate EGF-R signaling in human fetal intestine.
44-790G was used in immunocytochemistry to examine if EGF-R contributes to the hydrocortisone and indomethacin-induced intestinal perforations of preterm infants
|Kajanne R,Leppä S,Luukkainen P,Ustinov J,Thiel A,Ristimäki A,Miettinen PJ||Pediatric research (62:570)||2007|
|Mouse||Not Cited||Aplidin induces apoptosis in human cancer cells via glutathione depletion and sustained activation of the epidermal growth factor receptor, Src, JNK, and p38 MAPK.||Cuadrado A,Garcia-Fernandez LF,Gonzalez L,Suarez Y,Losada A,Alcaide V,Martinez T,Fernandez-Sousa JM,Sanchez-Puelles JM,Munoz A||The Journal of biological chemistry (278:241)||2003|