Using stand monoclonal attachment to plate, followed by 6x washes with PBST, the plate is blocked with 1.2% seablock followed with a 2%sucrose 4%OVO solution block. We have a high background with seablock, water or sample diluting buffer.
What are the remaining steps of your assay, particularly the enzyme conjugated reagents?
After the 45 minute block with the 2% sucrose and 4% PVP360, plates are dried and stored in foil pouches with dessicant and moisture cards.
The assay used serum with SurModics Assay Diluent. The Poly Alk Phos Conjugate uses the Surmodics AP Conjugate buffer.
Seablock uses salmon serum as a blocking agent. I am not that familiar with fish biology but I would imagine that fish serum would contain a lot of similar proteins, peptides, small molecules, etc. I would block with PBS with 1% BSA and 10% sucrose. That is what we use for blocking our antibody coated plates.
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