I am currently working with Mannheimia haemolytica. We make a membrane protein extraction with sodium salycilate (SS) and then we keep it with the SS at the same concentration (1 M). When kept at 4ºC all along the stability study we see that the protein pattern in SDS and WB keeps intact. Nevertheless, we loose signal in the ELISA in which we use the same antibody as in the WB.
So, something is lost but I don't know what it could be. Any suggestions?
Moreover, does anyone know or think of a way to improve membrane protein extraction stability from M.haemolytica at 4ºC (now it is stored in SS 1M solution)?
Just try it: Precipitate your proteins using acetone at 4ºC for overnight. Centrifuge it at 10K at 4ºC for 5 min. Air dry the sample and dissolve it in PBS and then check this protein by ELISA.
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