mick.partis at hri.ac.uk
Fri Nov 8 05:53:07 EST 1996
poseva at iata.csic.es (Eva Dupille) wrote:
>I have a protein problem precipitation. I did a TCA precipitation, 15 %
>for 1 h at 4 C, and I got, nothing at all.
>Sure that I have proteins, I get enzyme activity and with biorad, before
>precipitation, I have around 10 micrograms/ml.
>Is it a problem of temperature or incubation time ??
>Please, if you have some comments, let me know. That s boring to lose so
>many time in protein purification, to lose all the samples for a protein
>precipitation problem ..
>Thanks in advance, sorry for the approximative english.
You don't say why you want to precipitate your protein: if you are
running assays and just want to get rid of the protein then 15% TCA
should inactivate. If you need a precipitate then why not add a carrier
protein? Adding some serum albumin (or any cheap, pure protein) at about
1 mg/ml before adding your TCA and your protein should co-precipitate.
If you are precpitating prior to running SDS-PAGE, just make sure the MW
of the carrier protein is different from that of your protein.
mick.partis at hri.ac.uk Horticulture Research International
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