Help about TCA

alvarez at cica.es alvarez at cica.es
Thu Jun 5 01:17:01 EST 1997


Hello!

We have some problems with TCA precipitation of proteins. We use TCA 8%,
and after whashing the pellet with ethanol:diethylic ether 1:1, we leave
to dry at room temp. But, when we resuspend the pellet in loading
buffer, bromophenol blue turns yellow (pH below 5?), then we neutralize
with Tris base 1 M, pH 10.7. Before loading the sample in SDS-PAGE
minigel, we dilute it just to salt concentration 250 mM aprox. When the
gel is silver stained, the front is very dark, the lanes are full of
smearing. Control lanes (without TCA ect) run well.

Has someone any solution? Dialization is not a possibility.

Thanks

Jose



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