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Old 01-12-2004, 08:01 PM
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Default protein extraction from protplasts ?


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

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


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