I wanted to perform ELISA (sandwich elisa) on the protein pellet extracted from cell lysate. I would like to know in which buffer, the protein pellet should be resuspended?
Here is my protein extraction protocol from cell pellet to perform an Elisa test: -Resumption of cell pellets in the extraction buffer RIPA (200uL around for a pellet of a BP100 box) -Let the pellets rest in the ice for 2 hours with stirring -Centrifugation 20 min, 16000g, 4 ° C -Recover the supernatant -Preserve the supernatants at -20 ° C RIPA buffer: - Tris 50mM, pH 8 - 150mM NaCl - IGEPAL CA-630 1% - Deoxychelate sodium 0.5% - SDS 0.1% + cocktail inhibitors