Glycosylation of Bovine, Canine, Rabbit, and Swine Leukemia Inhibitory Factor

Kingfisher Biotech expresses recombinant proteins in yeast so they are potentially post-translationally modified. Bovine, canine, rabbit and swine Leukemia Inhibitory Factor (LIF) have calculated molecular weights of  19.8, 19.7, 19.7, and 19.7 kDa, respectively. The Kingfisher Biotech LIF recombinant proteins were 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 all four LIF recombinant proteins 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, canine, rabbit and swine LIF recombinant protein produced in yeast are glycosylated.

Lane	Sample	Treatment
1	Stds-PAGEgel Two-Color SDS Protein Ladder
2	Bovine LIF	Untreated
3	Bovine LIF	Denatured/Deglycosylated
4	Canine LIF	Untreated
5	Canine LIF	Denatured/Deglycosylated
6	Stds-PAGEgel Two-Color SDS Protein Ladder
7	Rabbit LIF	Untreated
8	Rabbit LIF	Denatured/Deglycosylated
9	Swine LIF	Untreated
10	Swine LIF	Denatured/Deglycosylated
11	Stds-PAGEgel Two-Color SDS Protein Ladder
12

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