Lane 1 MW markers; Lane 2 ab83676; Lane 3 ab83676 treated with PNGase F to remove potential N-linked glycans; Lane 4 ab83676 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. Slight drop in MW after treatment with glycosidase cocktail suggests presence of Olinked glycans. Additional bands in lane 3 and lane 4 are glycosidase enzymes.
A sample of ab83676 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 of the Chimera. Spots within the spot train were cut from the gel and identified as the Chimera by protein mass fingerprinting.
Post-translational modifications result in protein heterogeneity. The densitometry scan demonstrates the purified ab83676 exists in multiple isoforms, which differ according to their level of post-translational modification. Expression of these isoforms is highly significant for cell biology, as they more closely resemble the native human proteins. Triangle indicates theoretical pI and MW of the protein.