I use 5X 0.5M HEPES buffer and 100mM ZnSo4 Nacl buffer into 1x as column buffer with pH 7.4 and 1X column buffer with 0.5M maltose as an elution buffer. Please suggest how I can overcome this problem?
You can try this
Wash Buffer
1 mM EGTA, pH 8.0
Filter through a 0.22 μm filter before use.
Add the DTT, PMSF, and Na2S2O5 fresh before use.
200 mM KCl
50 mM Tris-HCl, pH 8.0
1 mM EDTA, pH 8.0
2 mM Sodium Metabisulfite (Na2S2O5)
0.5 mM DTT
1 mM PMSF
10% (v/v) Glycerol
Elution Buffer
1 mM EGTA, pH 8.0
Filter through a 0.22 μm filter before use.
Add the DTT, PMSF, and Na2S2O5 fresh before use.
200 mM KCl
50 mM Tris-HCl, pH 8.0
1 mM EDTA, pH 8.0
2 mM Na2S2O5
10 mM Maltose
0.5 mM DTT
1 mM PMSF
10% (v/v) Glycerol
Thank you Nilanjan Roy. I will try these buffers. But my protein requires a cation metal to be in functional form. Can I still use EDTA?
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