Immunofluorescence analysis of EGFR [pY992] was done on 70% confluent log phase HeLa cells treated with EGF (200 ng/mL for 10 min). The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with EGFR [pY992] Rabbit polyclonal Antibody (44786G) at 2µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows untreated cells. Panel f shows no primary antibody control. The images were captured at 20X magnification.
|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 992. 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 *|
|Flow Cytometry (Flow)||1:100-1:500|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||Assay Dependent|
* 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.
Molecular basis for multimerization in the activation of the epidermal growth factor receptor.
44-786G was used in flow cytometry to study the molecular basis for activation of the epidermal growth factor receptor by multimerization
|Huang Y,Bharill S,Karandur D,Peterson SM,Marita M,Shi X,Kaliszewski MJ,Smith AW,Isacoff EY,Kuriyan J||eLife (5:null)||2016|
Fyn inhibition rescues established memory and synapse loss in Alzheimer mice.
44-786G was used in immunohistochemistry - free floating to assess the Src family kinase inhibitor, AZD0530, for treatment of Alzheimer's disease.
|Kaufman AC,Salazar SV,Haas LT,Yang J,Kostylev MA,Jeng AT,Robinson SA,Gunther EC,van Dyck CH,Nygaard HB,Strittmatter SM||Annals of neurology (77:953)||2015|