We are running western blot for checking the ag-ab interactions.our proteins are from molecular weight 250 to 7 kda.But we are facing some technical issues with the transfer of proteins(i'm not sure if the problem is with transfer). Can anyone help us with any idea that can help with the complete transfer of the proteins to the pvdf membrane or a successful western blot experiment.
TIA
Hello Nisha Reghu
You may try the heat-mediated blotting method. It is a preferable method over the conventional protocol due to enhanced transfer of both high and low molecular weight proteins. The permeability of the gel is increased by heat, such that the proteins trapped in the polyacrylamide gel matrix can be easily transferred to the membrane, thus facilitating faster protein transfer.
You may want to refer to the articles attached below for more information.
Article Western Blot: Protein Transfer Overview
Article Western Blotting of High and Low Molecular Weight Proteins Using Heat
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