Hi all, I have been trying to detect several polypeptides by WB. And tricine-sds-page (16.5% or 20%) and glycine-sds page(20%) gel electrophoresis followed by Comassie blue staining showed no bands below 7.8kda marker, either bands in blotting(antibody works well). The attachment file is the Coomassie blue staining of a 20% tricine gel.
Any experience or communication will be appreciated!
What was the time and voltage? Since the Molecular weight of the proteins between 3 to 7 kDa is considered to be of low molecular weights, therefore, it is better to run the transfer buffer at a voltage between 75-90 for about 60-70 mins and not more than that for optimal results. Moreover, the temperature of the transfer buffer has much to do with improper transfer. The temperature should be low, optimally 4°C.
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