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