Components
Pre-coated 96-well strip microplate; Wash buffer; Stop solution; Assay diluent(s); Lyophilized standard; Biotinylated detection antibody; Streptavidin-conjugated HRP; TMB one-step substrate
Reagent Preparation
1.Distilled or deionized water 2.Precision pipettes to deliver 2 µL to 1 µL volumes 3.Adjustable 1-25 µL pipettes for reagent preparation 4.100 µL and 1 liter graduated cylinders 5.Tubes to prepare standard and sample dilutions 6.Absorbent paper 7.Microplate reader capable of measuring absorbance at 450nm 8.Log-log graph paper or computer and software for ELISA data analysis
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.
Storage
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Restrictions
For research use only. Intended for use by laboratory professionals.
Datasheet