About GST pulldown

Ning LI libra at ion.ac.cn
Sat Jun 1 23:50:14 EST 2002


Hi everybody
  I have several questions on GST pulldown assay. The GST protein has
been eluted from the resin with reduced glutathione. <Molecular
Cloning> suggests that the yield of GST protein could be estimated by
measuring the absorbance at 280nm. But many kits say that glutathione
absorbs at 280nm and masks the eluted protein. Which one is right? And
since glutathione could elute GST protein from the column, is it
needed to dialyze to remove the glutathione before incubating with the
cell lysate? Ohterwise the GST protein may not bind to the beads
again?


Ning Li
Institute of Neuroscience
Chinese Academy of Sciences




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