RIPA extraction works very well for western blot, but not with ELISA. I have read about acetic acid or TFA extraction protocols, however, they are not suitable to maintain protein stability. I am also worried about using columns since my peptide is in very low quantities, and using columns would lead to loss of my peptide. Would simple PBS extraction followed by lyophilzation to concentrate the Peptide work? or if I lyophilze RIPA extracts, would it be possible to get rid of salts that might be interfering with ELISA?
Hi, if your peptide is soluble, just homogenize in 0.1x PBS and centrifuge at min.30.000g for min. 15 min at 4°C, then take the supernatant, but avoid the fat on top of it, and measure. There are acidification protocols to extract more Protein but that depends on the Peptide you`re looking for.
If you want to get rido of salts in your extract you may try dialysis against distilled water overnight at 4°C.
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