Human Phospho-EGFR (Y845) Antibody

Catalog #: AF3394
4 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF3394
AF3394-SP

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

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Citations (4)

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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 either lyophilized or 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.

Scientific Data

Western Blot

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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.

Intracellular Staining by Flow Cytometry

Detection of Phospho-EGFR antibody in EGF-treated A431 Human Cell Line antibody by Flow Cytometry.

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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.

Immunocytochemistry

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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

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); 24329 (Rat); 102138724 (Cynomolgus Monkey)

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

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Product Datasheet

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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.

4 Citations: Showing 1 - 4
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RANK-C Expression Sensitizes ER-Negative, EGFR-Positive Breast Cancer Cells to EGFR-Tyrosine Kinase Inhibitors (TKIs)
Authors: C Sirinian, AD Papanastas, SE Degn, T Frantzi, C Aronis, D Chaniotis, T Makatsoris, A Koutras, HP Kalofonos
Genes, 2021-10-23;12(11):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
A nongenomic mechanism for “metalloestrogenic” effects of cadmium in human uterine leiomyoma cells through G protein-coupled estrogen receptor
Authors: Jingli Liu, Linda Yu, Lysandra Castro, Yitang Yan, Maria I. Sifre, Carl D. Bortner et al.
Archives of Toxicology
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, 2013-02-28;14(3):4783-92.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P
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., 2010-08-04;70(18):7304-14.
Species: Human
Sample Types: Tissue Homogenates
Applications: Western Blot