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protein precipitation - isolation of proteins (Jul/11/2011 )


I am doing TCA extraction of proteins for isolation of proteins .. i am using glass beads and ribolyzing it and then i am spinning down the sample adn removing the supernatant. i am trying to resusped the pellet in 250 micro ltrs of TCA LB + BME .. It is very difficult to re suspend them in the tca lb. can anybody suggest me regarding this ... when ido western blot with the sample. i am not getting thick signal.. its very weak.. i bioled the sample @ 95 deg for 5 min and spinned them down befre running sds page.


What I do not understand is why TCA is used for resuspension? For SDS-PAGE a Leammli buffer is to use

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