Deglycosylation of Bovine IL-6 (RP0014B)

Kingfisher Biotech expresses recombinant proteins in yeast so they are potentially post-translationally modified. Bovine IL-6 has a calculated molecular weight of 20.6 kDa. The Kingfisher Biotech Bovine IL-6 (RP0014B) recombinant protein was treated with deglycosylation enzymes using NEB kit Protein Deglycosylation Mix  (NEB Catalog No. P6039S) to determine if the protein produced in yeast is glycosylated. The results show that Bovine IL-6 shifts in molecular weight when treated with the deglycosylation enzymes; the smear of higher MW material appears to be reduced to a single band migrating with the 22/28 kDa doublet standards. The bands showing the molecular weight of the deglycosylation enzymes are also visible. This data suggests Kingfisher Biotech's Bovine IL-6 recombinant protein produced in yeast is glycosylated.

Lane	Sample
1	Standards-PAGE gel Two-Color SDS Protein Ladder
2	Bovine IL-6 KFB RP0014B-025 - untreated
3	Bovine IL-6 KFB RP0014B-025 - denatured
4	Bovine IL-6 KFB RP0014B-025 - denatured and deglycosylated
5	Standards-PAGE gel Two-Color SDS Protein Ladder

MW of Standards: 7.6, 12, 22/28 doublet, 33, 48, 71, 96, 142, 195 kDa. 
16% Acrylamide SDS-PAGE Gel (Expedeon)