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Human Phospho-EGFR (Y845) Antibody

Catalog #: AF3394
4 Citations Datasheet
Catalog # Availability Size / Price Qty
AF3394
AF3394-SP
Detection of Human Phospho-EGFR (Y845) by Western Blot.
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Product Details
Citations (4)
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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

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 Detection of Human Phospho-EGFR (Y845) antibody 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.

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

Immunocytochemistry Phospho-EGFR (Y845) antibody in A431 Human Cell Line by Immunocytochemistry (ICC). 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.

Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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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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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  1. 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
  2. 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
  3. 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
  4. 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

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