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Human/Mouse/Rat Phospho-EGFR (Y992) Cell-Based ELISA

CODE: CBEL-EGFR992-1

$371.00

RayBio® Cell-Based Human/Mouse/Rat EGFR (Tyr992) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.


Product Features

  • Site and signal pathway-specific
  • In vitro detection of adherent cell culture
  • No sample lysis is needed
  • Compatible with a standard ELISA plate reader
  • Faster results than with ELISA
  • Adaptable for high-throughput screening and drug discovery

Specificity

The antibodies provided in this kit recognizes human, mouse and rat EGFR phosphorylated at site Tyrosine-992 and total EGFR for comparison.

Target Protein Information

Synonyms
Epidermal growth factor receptor (EC 2.7.10.1) (Proto-oncogene c-ErbB-1) (Receptor tyrosine-protein kinase erbB-1)
Accession Number
Gene Names
EGFR ERBB ERBB1 HER1
Gene ID
1956

Species Detected

Human, Mouse, Rat

Application Notes

Kit Components
  • uncoated 96-well Microplate
  • Wash Buffer A
  • Wash Buffer B
  • Fixing Solution
  • Quenching Buffer
  • Blocking Buffer
  • Anti-phospho antibody
  • Anti-pan antibody
  • HRP-Conjugated Secondary Antibody
  • TMB One-Step Substrate
  • Stop Solution
  • Other Materials Required
    • Distilled or deionized water
    • 100 ml and 1 liter graduated cylinders
    • Tubes to prepare sample dilutions
    • Protease and Phosphatase inhibitors
    • Precision pipettes to deliver 2 µl to 1 ml volumes
    • Adjustable 1-25 ml pipettes for reagent preparation
    • Benchtop rocker or shaker
    • Microplate reader capable of measuring absorbance at 450 nm
    Protocol Outline
    1. Seed 10,000-30,000 cells into each well and incubate overnight.
    2. Apply various treatment, inhibitors or activators according to manufacture's instructions.
    3. Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
    4. Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
    5. Add 200 µl of Blocking Solution and incubate for 1 h at 37 °C.
    6. Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
    7. Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
    8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
    9. Incubate 30 min at RT.
    10. Add 50 µl of Stop Solution to each well.
    11. Read at 450 nm immediately.

    Storage/Stability

    The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

    Product Specifications

    • 1, 2, or 5 x 96-Well Microplate Kit

    Array/ELISA Format

    • HER/ErbB Family
    • Human
    • 96-well Microplate
    • Cell-based
    • Semi-Quantitative
    • Colorimetric
    • Adherent Cell Culture

    Note: If you are asking about pricing and you are outside of the US please contact your local RayBiotech distributor.

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    Usage

    This product is furnished for LABORATORY RESEARCH USE ONLY.
    Not for diagnostic or therapeutic use.

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