1D SDS-PAGE of ab84071 before and after treatment with glycosidases to remove oligosaccharides. Lane 1: ab84071 Lane 2: ab84071 treated with PNGase F to remove potential N-linked glycans Lane 3: ab84071 treated with a glycosidase cocktail to remove potential N- and O-linked glycans. 10 µg protein loaded per lane; Deep Purple™ stained. Drop in MW after treatment with PNGase F indicates presence of N-linked glycans. Subsequent tightening of band after treatment with glycosidase cocktail suggests presence of O-linked glycans. Additional bands in lane 2 and lane 3 are glycosidase enzymes.
A sample of ab84071 without carrier protein was reduced and alkylated and focused on a 3-10 IPG strip then run on a 4-20% Tris-HCl 2D gel. 40µg protein loaded per lane; Deep Purple™ stained. Spot train indicates presence of multiple isoforms. Spots within the spot train were cut from the gel and identified as IL2 Receptor alpha (Fc Chimera) by protein mass fingerprinting.
Densitometry of protein isoforms visualised by 2-DE. The densitometry scan demonstrates the purified human cell expressed protein exists in multiple isoforms, which differ according to their level of post-translational modification. The triangle indicates the theoretical MW and pI of the protein.