Purified anti-EGFR Antibody

Pricing & Availability
Clone
A19002A (See other available formats)
Other Names
Epidermal Growth Factor Receptor, Receptor Tyrosine-Protein Kinase ErbB-1, Erb-B2 Receptor Tyrosine Kinase 1, ERBB1, NISBD2, PIG61, MENA, Proto-oncogene c-ErbB-1
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
1_A19002A_PURE_EGFR_Antibody_1_111219_updated.png
Whole cell extracts (15 µg protein) from MOLT-4 (negative control), HeLa, PC-3, HaCaT, and A-431 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of purified anti-EGFR antibody (clone A19002A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-mouse IgG Antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904) was used as a loading control at a 1:50,000 dilution (lower). Lane M: Molecular weight marker.
  • 1_A19002A_PURE_EGFR_Antibody_1_111219_updated.png
    Whole cell extracts (15 µg protein) from MOLT-4 (negative control), HeLa, PC-3, HaCaT, and A-431 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of purified anti-EGFR antibody (clone A19002A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP Goat anti-mouse IgG Antibody (Cat. No. 405306) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH Antibody (Cat. No. 607904) was used as a loading control at a 1:50,000 dilution (lower). Lane M: Molecular weight marker.
  • 2_A19002A_PURE_EGFR_Antibody_2_103019.png
    A-431 cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with ice-cold methanol for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 5.0 µg/mL (1:100 dilution) of either Purified Mouse IgG1, κ Isotype Ctrl Antibody (Cat. No. 401402, panel A) or purified anti-EGFR antibody (panel B) for two hours at room temperature, followed by incubation with Alexa Fluor® 594 Goat anti-mouse IgG Antibody (Cat. No. 405326) at 2.0 µg/mL. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • 3_A19002A_PURE_EGFR_Antibody_6_111219.png
    Whole cell extracts (300 µg total protein) prepared from A-431 cells were immunoprecipitated overnight with 2.5 µg of Purified Mouse IgG1, κ Isotype Ctrl Antibody (Cat. No. 401401) or purified anti-EGFR antibody (clone A19002A). The resulting IP fractions and whole cell extract input (5%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of A18011A. Lane M: Molecular weight marker.
  • 4_A19002A_PURE_EGFR_Antibody_3_103019.png
    Formalin-fixed paraffin-embedded human breast cancer tissue slices were deparaffinized and rehydrated. Antigen retrieval was done with Tris-Buffered Saline 1X (1.0 M, pH 7.4) at 95°C for 40 minutes, washed with PBS/0.05% Tween 20 twice for five minutes, permeabilized with 0.5% Triton X-100 for ten minutes, and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the slices were stained with 5 µg/mL of purified anti-human EGFR (clone A19002A) at 4°C overnight, followed by 2.5 µg/mL of Alexa Fluor® 594 anti-mouse IgG (clone Poly4053) (red) for two hours at room temperature. Nuclei were counterstained with DAPI (green). The image was captured with a 10X objective.
  • 5_A19002A_PURE_EGFR_Antibody_4_103019.png
    Formalin-fixed paraffin-embedded human placenta tissue slices were deparaffinized and rehydrated. Antigen retrieval was done with Tris-Buffered Saline 1X (1.0 M, pH 7.4) at 95°C for 40 minutes, washed with PBS/0.05% Tween 20 twice for five minutes, permeabilized with 0.5% Triton X-100 for ten minutes, and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the slices were stained with 5 µg/mL of purified anti-human EGFR (clone A19002A) at 4°C overnight, followed by 2.5 µg/mL of Alexa Fluor® 594 anti-mouse IgG (clone Poly4053) (red) for two hours at room temperature. Nuclei were counterstained with DAPI (green). The image was captured with a 10X objective.
  • 6_A19002A_PURE_EGFR_Antibody_5_103019.png
    A-431 cells were stained intracellularly with purified anti-EGFR (clone A19002A) (filled histogram) or mouse IgG1, κ isotype control (open histogram), followed by PE anti-mouse IgG antibody.
Cat # Size Price Quantity Avail. Save
933901 25 µg £58
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933902 100 µg £147
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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 the protein. These phosphorylation events stabilize the EGFR kinase activation loop and lead 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

Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide from human EGFR
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IHC-P, ICFC, IP - Validated

Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.1 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 2.5 - 5 µg/mL is suggested. For flow cytometric staining, the suggested use of this reagent is ≤ 0.06 µg per million cells in 100 µL volume. For immunoprecipitation, the suggested use of this reagent is 2.5 µg/test. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone was tested for ICC using 4% PFA-fixed A431 cells permabilized with methanol or Triton X-100. While both permeabilization methods were compatible with the antibody, methanol enabled superior staining.

This clone is predicted to recognize isoform I based off of complete sequence homology of the immunizing peptide and the corresponding region of the isoform.

 

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
Cell Type
Endothelial cells, Epithelial cells
Biology Area
Cell Biology, Cell Cycle/DNA Replication, Immunology, Innate Immunity, Neuroscience, Synaptic Biology
Molecular Family
CD Molecules, Growth Factors
Antigen References
  1. Hunter T and Cooper JA. 1981. Cell. 24:741.
  2. Gill GN and Lazar CS. 1981. Nature. 293:305.
  3. Reynolds FH, et al. 1981. Nature. 292:259
  4. Zhou M, et al. 2013. Cancer Res. 23:7056-67
Gene ID
1956 View all products for this Gene ID
UniProt
View information about EGFR on UniProt.org

Related FAQs

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Go To Top Version: 1    Revision Date: 10/30/2019

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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.
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