aai140 at agora.ulaval.ca
Fri Nov 15 18:31:33 EST 1996
> > Hello,
> > 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.
You can try to precipitate your protein with following protocol:
0.15% deoxycholate (DOC)
To 1 ml of protein sample, add 0.1ml of DOC. Vortex and let it stand at RT
for 10 min
To the above add 0.1ml of 72% TCA. let it atand on ice for 30 min.
Spin the above at 17000 rpm for 30 min
Dissolve the pellet in SDS_loading buffer. If the sample is yellow (due to
TCA) add 1 µl of 250mM tris base
Voala, run the SDS_PAGE. Surely, you cannot visually see the 10µg of your
protein after TCA ppt. but after SDS_PAGE you should be able to see.
Try this protocol.
More information about the Proteins