Human Phospho-EGFR (Y845) Antibody
R&D Systems, part of Bio-Techne | Catalog # AF3394
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human
Cited:
Human
Applications
Validated:
CyTOF-ready, Immunocytochemistry, Immunohistochemistry, Intracellular Staining by Flow Cytometry, Western Blot
Cited:
Immunohistochemistry-Paraffin, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Product Specifications
Immunogen
Phosphopeptide containing human EGFR Y845 site
Specificity
Detects human EGFR when phosphorylated at Y845.
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human Phospho-EGFR (Y845) Antibody
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.
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.
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.Applications for Human Phospho-EGFR (Y845) Antibody
Application
Recommended Usage
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
Sample: Immersion fixed A431 human epithelial carcinoma cell line treated with pervanadate
Sample: Immersion fixed A431 human epithelial carcinoma cell line treated with pervanadate
Immunohistochemistry
5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human glioblastoma and sarcoma subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)
Sample: Immersion fixed paraffin-embedded sections of human glioblastoma and sarcoma subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Sample: A431 human epithelial carcinoma cell line treated with Recombinant Human EGF (Catalog # 236-EG), fixed with paraformaldehyde, and permeabilized with saponin
Sample: A431 human epithelial carcinoma cell line treated with Recombinant Human EGF (Catalog # 236-EG), fixed with paraformaldehyde, and permeabilized with saponin
Western Blot
0.5 µg/mL
Sample: A431 human epithelial carcinoma cell line treated with Recombinant Human EGF (Catalog # 236-EG)
Sample: A431 human epithelial carcinoma cell line treated with Recombinant Human EGF (Catalog # 236-EG)
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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.
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
Alternate Names
EGF R, ErbB, ErbB1, HER-1
Gene Symbol
EGFR
Additional EGFR Products
Product Documents for Human Phospho-EGFR (Y845) Antibody
Product Specific Notices for Human Phospho-EGFR (Y845) Antibody
For research use only
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