1D SDS-PAGE of ab84065 before and after treatment with glycosidases to remove oligosaccharides. Lane 1: ab84065 Lane 2: ab84065 treated with PNGase F to remove potential N-linked glycans Lane 3: ab84065 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 drop in MW after treatment with glycosidase cocktail indicates presence of O-linked glycans. Faint bands in lane 2 and lane 3 are glycosidase enzymes.
A sample of ab84065 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. 40 µg 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 gamma (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.