The antibodies provided in this kit recognizes human, mouse and rat EGFR phosphorylated at multiple sites (Tyrosine-phosphorylated EGFR, EGFR phosphorylated at site Tyrosine-1068, EGFR phosphorylated at site Tyrosine-845 and EGFR phosphorylated at site Tyrosine-992) and total EGFR for comparison.
Number of Targets Detected:
Compatible Sample Types:
Adherent Cell Culture
Method of Detection:
Two 96-Well Microplate Kit
Site and signal pathway-specific
In vitro detection of adherent cell culture
No sample lysis needed
Compatible with a standard ELISA plate reader
Faster results than with ELISA
Adaptable for high-throughput screening and drug discovery
Cell Culture Microplate
Wash Buffer A
Wash Buffer B
HRP-Conjugated Secondary Antibody
TMB One-Step Substrate
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
Seed 10,000-30,000 cells into each well and incubate overnight.
Apply various treatment, inhibitors or activators according to manufacture's instructions.
Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
Add 200 µl of Blocking Solution and incubate for 1 h at 37 °C.
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.
Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.