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protein extraction from protplasts ?
Hello
I need some information regarding extraction of protein from transfected tomato protoplasts. I am using 2x10(4) cells for PEG mediated transfection. I tried sonication, micropestle grinding, freeze-thawing,... but none of these seem to work. I tried to estimate protein concentration by BCA protein assay and Biorad protein assay. I got very low or negative readings. Does anybody know 1. how exactly protein is extracted from protoplasts 2. how many protoplasts are needed for extraction I need to do immunoprecipitation and RT-PCR experiments. Could somebody give me some insights into this. Thanks a lot in advance Roopa |
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