* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Product Specific Information
The M806B anti-EGF antibody (biotinylated conjugate of clone 3A8) has successfully been paired as the detection antibody in a sandwich ELISA with coating antibody M805 (Clone 1H11).
Typical dilutions for sandwich ELISA: Coat = 1-3 µg/ml and Detection = 0.125-0.5 µg/ml.
The protein encoded by EGF gene is a growth factor that stimulates cell growth, proliferation, and differentiation by binding to its receptor EGFR. Human EGF is a 6045-Da protein with 53 amino acid residues and three intramolecular disulfide bonds
EGF Antibody (M806B) in ELISA
ELISA with anti-EGF Monoclonal Antibody [3A8], Biotin conjugate (M806B)
Sandwich ELISA analysis of Human EGF was performed using the Thermo Scientific ELISA Kit (Product # EHEGF) by coating a blank 96-well microtiter plate with 100µl per well of Human EGF moncolonal antibody (Product # M805) in duplicate at 1, 3, 4, 5, 7, and 9µg/ml in DPBS (Product # 28374) and incubating for 12-18 hours at 4C. The plate was aspirated and blocked with 300µl per well of 4% BSA and 5% sucrose in DPBS for 1 hour at room temperature. Human EGF recombinant protein at 50µl per well was added in duplicate at 500, 250, 125, 62.5, 31.25, 15.625, 7.8125 and 0 pg/ml for 2 hours at room temperature along with 50µl of Human EGF biotinylated monoclonal antibody (Product # M806B) in all applicable wells at 0.2µg/ml for 2 hours at room temperature. The plate was washed with ELISA Wash Buffer (Product # N503) and incubated with 100µl per well of Streptavidin-HRP (Product # N504) in all test wells at 1:130,000 dilution for 1 hour at room temperature and then washed and incubated with 100µl per well of TMB substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was stopped with 0.16M sulfuric acid (Product # N600). Absorbances were read on a spectrophotometer at 450-550nm.
(This product is for In Vitro experimental use only. Not for resale without express authorization.)