Bovine Leukemia Inhibitory Factor (LIF) Polyclonal Antibody Testing

Using our bovine LIF recombinant protein as the antigen, polyclonal antibodies were produced in chickens and screened for recognition of both the glycosylated and deglycosylated forms of bovine, canine, rabbit and swine LIF recombinant proteins. (Previously it was demonstrated that the Kingfisher Biotech LIF recombinant proteins were glycosylated.) The Kingfisher Biotech LIF recombinant proteins were either treated with deglycosylation enzymes or left untreated and were subsequently screened using a Western Blot. The Western Blot showed that the anti-bovine LIF polyclonal antibody recognized both the glycosylated and the deglycosylated forms of bovine (lanes 2 & 3), canine (lanes 4 & 5), rabbit (lanes 6 & 7), and swine (lanes 7 & 8) LIF recombinant proteins.

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)

LIF Homology Across Species

Bovine, Canine Rabbit, and Swine Leukemia Inhibitory Factor Recombinant Proteins are available from Kingfisher Biotech. Leukemia inhibitory factor, or LIF, is a member of the IL-6 family. LIF affects cell growth by inhibiting differentiation. When LIF levels drop, the cells differentiate. 

Rabbit and Canine Leukemia Inhibitory Factor (LIF) Recombinant Proteins

Kingfisher Biotech's Rabbit and Canine Leukemia Inhibitory Factor (LIF) recombinant proteins can be used in cell culture, as an LIF ELISA Standard, and as a Western Blot Control. Leukemia inhibitory factor, or LIF, is a member of the IL-6 family. LIF affects cell growth by inhibiting differentiation. When LIF levels drop, the cells differentiate. Leukemia inhibitory factor derives its name from its ability to induce the terminal differentiation of myeloid leukemic cells, thus preventing their continued growth. Other properties attributed to LIF include: the growth promotion and cell differentiation of different types of target cells, influence on bone metabolism, cachexia, neural development, embryogenesis and inflammation. p53 regulated LIF has been shown to facilitate implantation in the mouse model and possibly in humans. Removal of LIF pushes stem cells toward differentiation, but they retain their proliferative potential or pluripotency. LIF is typically added to stem cell culture medium to reduce spontaneous differentiation.

Kingfisher Biotech's LIF recombinant proteins are intended for use in animal model and animal health research. Our portfolio includes bovine, canine, and rabbit Leukemia Inhibitory Factor (LIF) recombinant proteins. We will be adding Swine Leukemia Inhibitory Factor (LIF) recombinant protein soon.