Question

What's the purpose of these in protein Purification of 6xHIS-Ubiquitin specific protease and GST EBNA1

Sample Purpose of the sample in this experiment Full name GSTE-NI GSTE non indced protedse Lysae 3. GSTE Lys GSTE LYsaHe 4. GSTR 5. GSTE R GST Rest GSTE Resin 7. GST PD-UP PVLL DO WN GST 8. GSTE PD-UP GSTE 9. GST PD-W 10. GSTE PD-WGSTE Pull hn W ELUTIUN ELUTION 11. GST PD-Elt 6ST PMIL pow 12. GSTE PD-Elt 6STE PALL DOw

Answer 1

6xHis-USP7 tag used to purify the protein by using Ni2+ ions kept in the column.

GST high affinity to GSH (Glutathione).

EBNA1 BINDS WITH GST (GSTE)

GST and GSTE was passed over the column and attached to the GSH in the column. The 6xHis USP7 was flowed through the column. GST has no affinity for USP7 however, GSTE does have. Therefore, GSTE would attach to the USP7.