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EGFR In-Cell ELISA Kits


Microplate assays to measure total and phospho-EGFR in whole cells.

Pierce EGFR In-Cell ELISA Kits

Thermo Scientific Pierce EGFR In-Cell ELISA Kits are colorimetric and fluorescent microplate assays optimized to measure total and phosphorylated epidermal growth factor receptor (phospho-EGFR) in whole cells.

These ELISA kits for whole cells include a mouse monoclonal anti-EGFR antibody, rabbit monoclonal anti-phospho-(Y1173) EGFR antibody, detection reagents and all buffers required for either colorimetric or near infrared detection. The EGFR In-Cell ELISA Kits enable simultaneous detection of both total and phospho-(Y1173) EGFR.

Highlights:

  • Measures total and phosphorylated epidermal growth factor receptor (EGFR) in whole cells using 96-well or 384-well microplates
  • Two reliable detection formats available:
    • colorimetric detection kit for use with standard microplate readers
    • near infrared fluorescent detection kit for use with IR imaging system
  • Whole-cell detection requires no cell lysis, protein extraction or special sample preparation
  • Provides better data and higher throughput than quantitative Western blotting
  • Less costly (one well vs. one lane) than Western blotting

Product Details:

Comparision of Near IR and Colorimetric Detection Kit Results Thermo Scientific Pierce EGFR In-Cell ELISA Kits give similar results with near IR and colorimetric detection methods. Both the near infrared and colorimetric detection formats for the EGFR In-Cell ELISA Kits show similar fold-induction in EGF stimulated A431. Z-factor (0.82 and 0.80, respectively) indicate comparable and robust performance between the near IR and colorimetric formats.

Near IR detection of EGFR near IR detection of EGFR with PD
Graph EGFR data with overlay of near IR and colorimetric detection methods

Graph EGFR data with overlay of near IR and colorimetric detection methods

The In-Cell ELISA Kit provides a fast and reliable format for determination of EGFR protein levels, activation and inhibition. Evaluation of dose-dependent activation and inhibition of EGFR in A431 cells with EGF and PD169393. Panels A and B show an increase in phospho-EGFR with increasing doses of EGF. The EGF-induced phosphorylation of EGFR is inhibited with increasing concentrations of PD169393 (panel C, rows C and D), whereas total EGFR does not significantly change with either treatment. No significant staining is seen in the absence of primary antibodies (panel A, column 1 and panel C, column 12). Panels B and D show that the fluorescence (RIU, left Y-axis) values obtained with the near infrared detection method can be overlaid on top of the data obtained with the colorimetric detection method (right Y-axis).

Overview of EGFR and phosphorylation of EGFR:

Epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein and receptor tyrosine kinase and plays a vital role in the regulating many of the main cellular signal transduction pathways. Binding of EGFR by ligands like epidermal growth factor (EGF) and transforming growth factor (TGF) alpha cause conformational changes and activation of EGFR tyrosine kinase activity through receptor dimerization and autophosphorylation. Tyr 1068, Tyr 1148, and Tyr 1173 are the major sites of EGF-dependent autophosphorylation. The phosphotyrosines on EGFR mediate signal transduction through interaction with signaling proteins like of Grb2 and SHC. Phosphorylation of Tyr 1173 on EGFR mainly regulates signaling through the MAP kinase pathway. The EGFR In-Cell ELISA Kit enables simultaneous detection of both total and phospho-(Y1173) EGFR.

Related Resources:

Overview and Menu of Target-Specific In-Cell ELISA Kits
Dylight Fluor Spectral Properties (Ex/Em)

Related Products:

ELISA reagent reservoirs and sealing tape
Secondary Antibodies and Conjugates For ELISAs and Western Blotting
DyLight Fluor Conjugated Secondary Antibodies and Biotin Binding Proteins

Notice:

This product is for In Vitro experimental use only. Not for resale without express authorization.


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
62205 EGFR Colorimetric In-Cell ELISA Kit
Formulation: Multi-component kit
Sufficient For: 96 microplate wells of overnight-cultured cells

Kit Contents:
Blocking Buffer, 50mL
20X Tris Buffered Saline, 50mL
Surfact-Amps 20 (10% Detergent), 10mL
Surfact-Amps X-100 (10% Detergent), 10mL
HRP Conjugate, 0.14mL
TMB Substrate, 58mL
TMB Stop Solution, 55mL
Janus Green Whole-Cell Stain, 50mL
Elution Buffer, 2 x 30mL
Thin Plate Seal Assembly, 8 each
Anti-Phospho EGFR (Y1173) Antibody
Anti-EGFR Antibody
96-test kit Product Instructions for product #62205 EGFR Colorimetric In-Cell ELISA Kit MSDS for product #62205 EGFR Colorimetric In-Cell ELISA Kit $380.00

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62210 EGFR Near Infrared In-Cell ELISA Kit
Formulation: Multi-component kit
Sufficient For: 96 microplate wells of overnight-cultured cells

Kit Contents:
Blocking Buffer, 100mL
20X Tris Buffered Saline, 50mL
Surfact-Amps 20 (10% Detergent), 10mL
Surfact-Amps X-100 (10% Detergent), 10mL
DyLight 680 Goat Anti-Mouse IgG (H+L), 55µL
DyLight 800 Goat Anti-Rabbit IgG (H+L), 55µL
Thin Plate Seal Assembly, 8 each
Anti-Phospho EGFR (Y1173) Antibody
Anti-EGFR Antibody
96-test kit Product Instructions for product #62210 EGFR Near Infrared In-Cell ELISA Kit MSDS for product #62210 EGFR Near Infrared In-Cell ELISA Kit $380.00

Add to cart

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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