Sean Lauber:ELISA Development: Difference between revisions
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(New page: First you need to determine what reagents you will use for your ELISA. The protocol I use is based off of a generic R&D ELISA. The primary antibody binds to the plate (this should be monoc...) |
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Revision as of 08:45, 11 July 2012
First you need to determine what reagents you will use for your ELISA. The protocol I use is based off of a generic R&D ELISA. The primary antibody binds to the plate (this should be monoclonal), the sample binds to the primary, then the secondary binds to the sample (this should be be biotinylated), then strep-HRP binds to biotin and the HRP is used for color development (see any R&D ELISA for the complete protocol).
Next you need to determine what concentration of primary and secondary will work best for giving good color development for a range of sample concentrations. I choose the range 0.25 - 8 μg/mL for the primary, and 50 - 400 ng/ml for the secondary
