Human Phospho-EGFR (Y845) Antibody

(2 citations)   
  • 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. 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.
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. 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.
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/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
Related Research Areas
Citations:

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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Species
Applications
Sample Type
  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 Type: Whole Tissue
    Application: 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 Type: Tissue Homogenates
    Application: WB
Cell and Tissue Staining Kits
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Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit

CTS005 3
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Anti-Rabbit HRP-AEC Cell & Tissue Staining Kit

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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 4
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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Isotype Controls
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Normal Rabbit IgG Control

Ctrl AB-105-C 43  
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Normal Rabbit IgG Control

Ctrl, CyTOF-ready MAB1050  
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Secondary Antibodies
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Rabbit IgG HRP-conjugated Antibody

WB HAF008 23  
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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL004 10
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Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL005 2
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Rabbit IgG APC-conjugated Antibody

Flow F0111 1  
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Rabbit IgG PE-conjugated Antibody

Flow F0110 2  
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Rabbit IgG Fluorescein-conjugated Antibody

Flow F0112 5  
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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL006 4
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Goat Anti-Rabbit IgG Biotinylated Antibody

WB BAF008 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rabbit IgG Antibody

WB AF008 1
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Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC003  
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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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