heating of samples during plant protein extraction - (Nov/08/2006 )
Hi, I have recently carried out an experiment on the optimization of a plant total protein extraction protocol. I have found that heating the vegetable sample at 70 degrees C for 4 minutes prior to loading into an SDS-PAGE gel actually improves results. i.e, more distinct bands are produced. Does anyone have an explanation for this?
this happens because the proteins are denatured by the heat, and stay denatured in the SDS containing loading buffer and gel.
If you don't heat (most people boil for 3-5 min) your samples, the poteins can associate into concatemers and through other interactions. which won't be separated by the electrophoresis.
thanks for your help. Do you happen to know of any online references which explains about this? I'm doing this for my final year project, so I need to have some evidence of where I get my info from.