详细说明
Species Reactivity
Human
Specificity
Detects human EGFR when phosphorylated at Y1173 in Western blots.
Source
Polyclonal Rabbit IgG
Purification
Antigen Affinity-purified
Immunogen
Phosphopeptide containing human EGFR Y1173 site
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated
Applications
Recommended
ConcentrationSample
Western Blot
0.2 µg/mL
See below
Simple Western
2 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Immunocytochemistry
5-15 µg/mL
See below
Intracellular Staining by Flow Cytometry
2.5 µg/10 6 cells
See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. are available in the Technical Information section on our website.
Data Examples
Intracellular Staining by Flow Cytometry | Detection of Phospho-EGFR (Y1173) in A431 Human Cell Line by Flow Cytometry. A431 human epithelial carcinoma cells were untreated (open histogram), or treated for 5 minutes with 100 ng/mL Recombinant Human EGF (Catalog # ; filled histogram) then stained with Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1095), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # ). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. |
Western Blot | Detection of Human Phospho-EGFR (Y1173) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (‑) or treated (+) with 100 μM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody, followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # ). A specific band was detected for Phospho-EGFR (Y1173) at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using . |
Immunocytochemistry | Phospho-EGFR (Y1173) in A431 Human Cell Line. EGFR phosphorylated at Y1173 was detected in immersion fixed A431 human epithelial carcinoma cell line untreated (lower panel) or treated (upper panel) with pervanadate using Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # ) and counterstained with DAPI (blue). View our protocol for . |
Immunohistochemistry | Phospho-EGFR (Y1173) in Mouse Embryo. EGFR phosphorylated at Y1173 was detected in immersion fixed frozen sections of mouse embryo using Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # ) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for . |
Simple Western | Detection of Human Phospho-EGFR (Y1173) by Simple WesternTM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human EGF (Catalog # ) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-EGFR (Y1173) at approximately 265 kDa (as indicated) using 2 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system. |
Preparation and Storage
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: EGFR
Epidermal growth factor receptor (EGFR, also known as ErbB1 and HER1) is the founding member of the ErbB family of receptor tyrosine kinases. Ligand binding induces receptor dimerization and autophosphorylation on multiple tyrosine residues. Phosphorylation of Y1173 creates a binding site for the protein tyrosine phosphatase SHP-1, leading to attenuation of receptor signaling.
Long Name:
Epidermal Growth Factor Receptor
Entrez Gene IDs:
1956 (Human); 13649 (Mouse)
Alternate Names:
avian erythroblastic leukemia viral (v-erb-b) oncogene homolog; cell growth inhibiting protein 40; cell proliferation-inducing protein 61; EC 2.7.10; EC 2.7.10.1; EGF R; EGFR; epidermal growth factor receptor (avian erythroblastic leukemia viral (v-erb-b)oncogene homolog); epidermal growth factor receptor; ErbB; ErbB1; ERBB1PIG61; HER1; HER-1; mENA; Proto-oncogene c-ErbB-1; Receptor tyrosine-protein kinase erbB-1