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Purified anti-EGFR Phospho (Tyr845) Antibody

Pricing & Availability
Clone
Poly21A03 (See other available formats)
Regulatory Status
RUO
Other Names
Epidermal Growth Factor Receptor, Receptor Tyrosine-Protein Kinase ErbB-1, Erb-B2 Receptor Tyrosine Kinase 1, ERBB1, NISBD2, PIG61, MENA
Isotype
Rabbit Polyclonal IgG
Ave. Rating
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Product Citations
publications
Poly21A03_PURE_EGFR_Antibody_020122
Whole cell extracts (15 µg protein) from untreated (-) and recombinant human EGF-treated (+) (Cat. No. 585506) A-431 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a 1:2000 dilution of purified anti-EGFR Phospho (Tyr845) (clone Poly21A03) for two hours at room temperature. Proteins were visualized by chemiluminescence detection using HRP donkey anti-rabbit IgG (Cat. No. 406401) at a 1:3000 dilution. Total EGFR loading was confirmed using purified anti-EGFR (clone A19002A) (Cat. No. 933902) at 0.1 µg/mL. Western-Ready™ ECL Substrate Plus Kit (Cat. No. 426317) was used as a detection agent. Lane M: Molecular weight marker
  • Poly21A03_PURE_EGFR_Antibody_020122
    Whole cell extracts (15 µg protein) from untreated (-) and recombinant human EGF-treated (+) (Cat. No. 585506) A-431 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a 1:2000 dilution of purified anti-EGFR Phospho (Tyr845) (clone Poly21A03) for two hours at room temperature. Proteins were visualized by chemiluminescence detection using HRP donkey anti-rabbit IgG (Cat. No. 406401) at a 1:3000 dilution. Total EGFR loading was confirmed using purified anti-EGFR (clone A19002A) (Cat. No. 933902) at 0.1 µg/mL. Western-Ready™ ECL Substrate Plus Kit (Cat. No. 426317) was used as a detection agent. Lane M: Molecular weight marker
  • Poly21A03_PURE_EGFR_Antibody_2_020122
    ICC staining of purified anti-EGFR Phospho (Tyr845) (clone Poly21A03) on serum-starved A-431 cells untreated (negative control, panel A) and recombinant human EGF-treated A-431 (Cat. No. 585506) (positive control, panel B). The cells were fixed with 4% PFA and permeabilized with 100% methanol, and blocked with 5% FBS for 1 hour at room temperature. The cells were then stained with a 1:40 dilution of the primary antibody, followed by incubation with 2.5 µg/mL of Alexa Fluor® 594 donkey anti-rabbit IgG antibody (Cat. No. 406418) for 1 hour at room temperature. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
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602251 50 µL £201
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602252 200 µL £474
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Description

Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase that links extracellular mitogenic ligand binding to complex downstream signaling cascades. Initial ligand binding results in receptor oligomerization and autophosphorylation of multiple tyrosine residues within cytosolic domains of the protein. Phosphorylation at tyrosine 845, which is found in the kinase activation loop, stabilizes the active conformation and leads to the recruitment of adaptor proteins and other downstream effectors. EGFR stimulation leads to activation of RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCγ-PKC and STAT signaling cascades, and constitutive activation of the receptor promotes tumorigenesis in multiple cancers.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Antibody Type
Polyclonal
Host Species
Rabbit
Immunogen
Synthetic peptide corresponding to human EGFR phosphorylated at tyrosine 845
Formulation
PBS/0.09% NaN3/50% glycerol
Preparation
The antibody was purified by affinity chromatography.
Concentration
Lot-specific (to obtain lot-specific concentration, please enter the lot number in our Concentration and Expiration Lookup or Certificate of Analysis online tools.)
Storage & Handling
Upon receipt, store undiluted at -20°C.
Application

WB - Quality tested
ICC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested dilution of this reagent is 1:2000. For immunocytochemistry, a dilution of 1:40 is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

During product development testing for western blot, clone Poly21A03 displayed high sensitivity. We recommend using Western-Ready™ ECL Substrate Plus Kit (Cat. No. 426317) for detection, and using shorter primary incubation times.

For ICC, we recommend using 4% PFA-fixed cells permeabilized with Triton X-100 or methanol.

RRID
AB_2910475 (BioLegend Cat. No. 602251)
AB_2910475 (BioLegend Cat. No. 602252)

Antigen Details

Structure
EGFR is a 1,210 amino acid protein with a predicted molecular weight of 134 kD; the processed form is 1,186 amino acids in length.
Distribution

Plasma membrane/Ubiquitously expressed

Function
EGFR signaling, cell signaling
Ligand/Receptor
EGF
Antigen References
  1. Hunter T and Cooper JA. 1981. Cell. 24:741-52.
  2. Gill GN and Lazar CS. 1981. Nature. 293:305-7.
  3. Reynolds FH, et al. 1981. Nature. 292:259-62.
  4. Biscardi JS, et al. 1999. J Biol Chem. 274:8335-42.
  5. Zhou M, et al. 2013. Cancer Res. 23:7056-67-67.
Gene ID
1956 View all products for this Gene ID
UniProt
View information about EGFR Phospho Tyr845 on UniProt.org

Related FAQs

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Go To Top Version: 2    Revision Date: 04/01/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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