I want to detect the specific protein (Adiponectin) in FBS. This protein has great quantity in circulation and serum. Adiponectin have multi-form in circulation and serum. If I use a common antibody (most of antibody can cross-reactive with mouse and human, not bovine). I think It's very difficult to evaluate what kinds of oligomer in FBS and the quantity. Have any better idea for this experiment?
There is the blast comparison of human (gi|27807433|ref|NP_777167.1|) and bovine (gi|4757760|ref|NP_004788.1|) Adiponectin:
Identities 185/213(87%);
Positives 196/213(92%)
it means for me exactly the same protein according to amino-acid seq. The reason why you haven't found the info about its on producer page is simply, they havent' tested it.
I' would find any polyclonal AB raised against known as long as possible seq of Adiponectin, compare it with bovine. When 70% of AA is identical in within the antigene used it is more than enough to be sure (95% ;) will work with your bovine ver. Remember to order the IIAB against the heavy chain only (52kDa) since the forms of light chains are around your protein 20-30kDa is according to EXPASY 26.133 kDa the calculated mass of Adiponectin.
What to use instead of bovine milk or BSA are the thypical blocking agents you can expect Adiponectin there?
I can answer only theoretical bla-bla about substances i have heard or the company-dealers are describing as better than anythig before milk and BSA (but sitill 99% of lab people are using them)...
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