Components
Pre-coated 96-well strip microplate; Wash buffer; Anti-phospho antibody; Anti-pan antibody; HRP-conjugated secondary antibody; Streptavidin-conjugated HRP; Assay diluent; TMB one-step substrate; Stop solution; Lysis buffer; Positive control sample
Reagent Preparation
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
Assay Procedure
1.Prepare all reagents, samples and standards as instructed in the manual; 2.Add 100 µL of standard or sample to each well; 3.Incubate 2.5 h at RT or O/N at 4°C; 4.Add 100 µL of prepared biotin antibody to each well; 5.Incubate 1 h at RT; 6.Add 100 µL of prepared Streptavidin solution to each well; 7.Incubate 45 min 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.
Target/Description
The epidermal growth factor receptor (EGFR) is a transmembrane protein that is a receptor for members of the epidermal growth factor family (EGF family) of extracellular protein ligands.
Restrictions
For research use only. Intended for use by laboratory professionals.
Datasheet
