Human Phospho-EGFR (Y845) Antibody

Catalog # Availability Size / Price Qty
AF3394
AF3394-SP
Detection of Phospho-EGFR in EGF-treated A431 Human Cell Line by Flow Cytometry.
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Product Details
Citations (2)
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Human Phospho-EGFR (Y845) Antibody Summary

Species Reactivity
Human
Specificity
Detects human EGFR when phosphorylated at Y845.
Source
Polyclonal Rabbit IgG
Purification
Antigen Affinity-purified
Immunogen
Phosphopeptide containing human EGFR Y845 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 Concentration
Sample
Western Blot
0.5 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
Immersion fixed paraffin-embedded sections of human glioblastoma and sarcoma subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)
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/106 cells
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Intracellular Staining by Flow Cytometry Detection of Phospho-EGFR in EGF-treated A431 Human Cell Line by Flow Cytometry. View Larger

Detection of Phospho-EGFR in EGF-treated A431 Human Cell Line by Flow Cytometry. A431 human epithelial carcinoma cell line was untreated (open histogram), or treated for 5 minutes with 100 ng/mL Recombinant Human EGF (Catalog # 236-EG; filled histogram) then stained with Rabbit Anti-Human Phospho-EGFR (Y845) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF3394), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). Rabbit IgG (Catalog # AB‑105‑C, data not shown) was used as a control antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Western Blot Detection of Human Phospho-EGFR (Y845) by Western Blot. View Larger

Detection of Human Phospho-EGFR (Y845) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y845) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3394), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-EGFR (Y845) at approximately 175 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry Phospho-EGFR (Y845) in A431 Human Cell Line. View Larger

Phospho-EGFR (Y845) in A431 Human Cell Line. EGFR phosphorylated at Y845 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 (Y845) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3394) 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 # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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/HER1) is the founding member of the EGFR family of receptor tyrosine kinases. Ligand binding induces receptor dimerization and autophosphorylation on multiple tyrosine residues. Phosphorylation of tyrosine 845 is associated with regulation of receptor function and tumor progression.

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

Product Datasheets

Citations for Human Phospho-EGFR (Y845) Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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  1. Analysis of Epithelial Growth Factor-Receptor (EGFR) Phosphorylation in Uterine Smooth Muscle Tumors: Correlation to Mucin-1 and Galectin-3 Expression.
    Authors: Weissenbacher, Tobias, Vrekoussis, Thomas, Roeder, David, Makrigiannakis, Antonis, Mayr, Doris, Ditsch, Nina, Friese, Klaus, Jeschke, Udo, Dian, Darius
    Int J Mol Sci, 2013;14(3):4783-92.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC Paraffin-embedded
  2. Coactivated platelet-derived growth factor receptor alpha and epidermal growth factor receptor are potential therapeutic targets in intimal sarcoma.
    Authors: Dewaele B, Floris G, Finalet-Ferreiro J, Fletcher CD, Coindre JM, Guillou L, Hogendoorn PC, Wozniak A, Vanspauwen V, Schoffski P, Marynen P, Vandenberghe P, Sciot R, Debiec-Rychter M
    Cancer Res., 2010;70(18):7304-14.
    Species: Human
    Sample Types: Tissue Homogenates
    Applications: WB

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Cell and Tissue Staining Kits

Immunohistochemistry Reagents

Isotype Controls

Normal Rabbit IgG Control

Ctrl,CyTOF-ready

Reconstitution Buffers

Secondary Antibodies

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