Hi all, My protein is relatively pure except the dimer. i used 10 mM TCEP to reduce it (directly added 1 M TCEP to make final volume of 10mM and kept at room temperature for 10 mins] then i dialysed the protein sample to remove TCEP [dialysis buffer is Na acetate pH 4.0].
On performing SDS PAGE analysis after dialysis of the protein we are getting no dimer band, only band of our protein is observed and same is the case with UV reading there is no change in it. But the HPLC analysis of the protein shows two peaks instead of one peak as observed before TCEP treatment. what can be the reason for this. Kindly guide. i need the protein to formulate and conduct stability studies on the sample. the protein we obtain after IEX is pure except the dimer and i do not want to go for SEC as it greatly reduces protein content and also is quite time consuming. any light on what is happening will bevery useful. thanks and regards
