Components
Pre-coated 96-well strip microplate; Wash buffer; Anti-phospho antibody; HRP-conjugated secondary antibody; 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
Prepare all reagents and samples as instructed in the manual; Add 100 µL of sample or positive control to each well; Incubate 2.5 h at RT or O/N at 4 °C; Add 100 µL of prepared primary antibody to each well; Incubate 1 h at RT; Add 100 µL of prepared 1X HRP-Streptavidin to each well; Incubate 1 h at RT; Add 100 µL of TMB One-Step Substrate Reagent to each well; Incubate 30 min at RT; Add 50 µL of Stop Solution to each well; Read at 450 nm immediately.
Target/Description
Receptor tyrosine-protein kinase erbB-3, also known as HER3 (human epidermal growth factor receptor 3), is a membrane bound protein that in humans is encoded by the ERBB3 gene.
Restrictions
For research use only. Intended for use by laboratory professionals.
Datasheet
