Recombinant Equine IL-1RA Protein

Proteins|Recombinant Proteins|Animal Proteins

Blue ice

Recombinant protein product has a calculated molecular mass of 17 kDa. Due to the abundant glycosylation, it migrates as multiple large protein bands (17-40 kDa) in SDS-PAGE under DTT, beta-mercaptoethanol reducing conditions. After deglycosylation under denature condition, the protein presented as one single 17 kDa band. See deglycosylation analysis in SDS-PAGE image.

SDS-PAGE under reducing conditions and visualized by Coomassie blue staining

12352202

His26-Gln177

His-tag affinity purification by immobilized metal ion affinity chromatography (IMAC)

Figure 1. Deglycosylation of purified recombinant proteins. The same amount of purified proteins were untreated (Lane 2) or treated with protein deglycosylation enzymes under native (Lane 3) or reducing (Lane 4) conditions. Deglycosylation treatment resulted in a mobility shift of the protein to produce one major band at the expected size, thus indicating that the untreated recombinant protein (Lane 2) was glycosylated.Lane 1: protein standard ladder (kDa).Lane 2: untreated protein under reducing conditions.Lane 3: treated protein with deglycosylation enzymes under native conditions.Lane 4: treated protein with deglycosylation enzymes under denature conditions.Lane 5: deglycosylation mixture only without target proteins.