Hi,

I work with membrane protein and SDSL EPR. I use MTSL for labeling. My problem is I get very low yield after rebinding (from 5 mg/ml to 0.8 mg/ml) which is not enough to downstream work. I used Ni-NTA affinity column for purification.

I follow following procedure after first elution.

1-Elute protein in LMPG micelle

2- Label the protein using MTSL

3- Wash with phosphate buffer in LMPG

4- rebinding the labelled protein using Ni-NTA resin in LMPG

5-Elute the final protein in SDS micelle (get very low yield here)

Could any one help me to get more yield in my second elution?

Dhilhani