I have 2 proteins-one is 19KD and other is 5KD. I tried detecting both of them through western blot using 20% SDS gel. I transferred them into 0.45 PDVF membrane for 100mins, I could detect the 19KD protein but I couldn't get the 5KD protein.
Any suggestions will be appreciated.
Thanks in advance
Because u have to take 2-5% of SDS gel for low mol.wt.. pore size is also small
Use a lower pore size membrane, such as a 0.2 µm PVDF or nitrocellulose membrane, which can better retain small proteins. Optimize transfer conditions: Decrease the methanol concentration in the transfer buffer (to 10-15%) to improve protein transfer efficiency, especially for low molecular weight proteins. Sometimes shorter transfer times (e.g., 30-60 minutes) can help prevent small proteins from passing through the membrane.
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