Hi,
This is going to be an off topic question concerning this community. I have a protein 6XHis tagged. When retrieved from the Ni-NTA column with imidazole found to be degraded, appears like a deep band with other bands (touching each other below the main band) in SDS PAGE. The protein is a DNA binding (pI~ 10) and Tris-Hcl buffer is used with 10% glycerol and 300mM NaCl. for purification. Does Phosphate buffer do any help in stopping the degradation.
All suggestions are welcome. Thank you for your reply in advance.
Sincerely
Debajyoti Dutta