Harvesting protocol for 35S-protein on monolayer cells in 96well format

S. McKay S.Mckay at sct.gu.edu.au
Tue Dec 15 18:47:12 EST 1998


I am having problems harvesting monolayer cell lysates from a 96well plate.
Previously I have used trypsin but as I am currently undertaking binding
studies I fear that this will cause the disocciation of my protein from the
cell membrane.

My main problem is trying to find something which is compatible with both
scintillant (I'm using 35S labelling of my protein) but that won't dissolve
plastic (ie doesn't have an organic base to it).  I've tried 1M NaOH/0.5%
TritonX-100, 3% TritonX-100 and NCS tissue solubliser all to no avail.
Have considered SDS but haven't had the chance to try this out yet.  Was
wondering what other people have been using.

Thanks for any suggestions.


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