I'm trying to refold a protein ~42 kDa and the pI ~5.9(Fusion His-Ubiq). The protein itself is about 32 kDa.
I did a screen test for rfolding and at the end the fusion protein was cleaved and they seemed good on SDS gel.
Iused Gel filtration S-75 column to separate my protein from the tag. My protein disappeared on the column.
My buffer was 20mM Tris-HCl, pH8.0, 150 mM NaCl and 1mM DTT.
any idea or suggestion about my procedure or another protocols will be highly appreciated
Refolded protein on Gel Filtration
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