Description
General description
Recombinant B18R from vaccinia virus is expressed in human HEK 293 cells as an N-terminally his-tagged glycoprotein with a calculated molecular mass of 38.4 kDa. The DTT-reduced protein migrates as a ~50 kDa polypeptide on SDS-PAGE due to glycosylation. The non-reduced protein migrates as a ~46 kDa polypeptide on SDS-PAGE due to internal cystines. This protein is manufactured in human cells, with no serum. The human cells expression system allows human-like glycosylation and folding, and often supports better stability of the protein in culture.
Application
B18R protein can be used with the Simplicon RNA Reprogramming Technology (Catalog No. SCR550) to create “foot-print” free human iPS cells at the highest efficiency without unwanted RNA degradation caused by cellular IFN responses.
Biochem/physiol Actions
B18R is a vaccinia virus gene product that acts as a decoy receptor for Type I Interferons (IFNs). B18R binds type I IFNs more avidly than their receptor. This protein has been shown to inhibit all IFN-α subtypes and IFN-β, but not type II or III IFNs. B18R enables increased cell viability during RNA transfection protocols designed to convert human somatic donor cells into iPSCs via direct delivery of synthetic mRNA′s. As a type I IFN receptor, B18R has a broad species specificity. It has significant regions of homology with the alpha subunits of the mouse, human, and bovine type I IFN receptors. Binding of soluble recombinant B18R to cells protected the cultures from IFN and allowed vaccinia virus replication. B18R can be used with the Simplicon RNA Reprogramming Technology (Cat. No. SCR550) to create “foot-print” free human iPS cells at the highest efficiency without unwanted RNA degradation caused by cellular IFN responses.
Physical form
This product is supplied as a 50 μg liquid solution, with 10 mM phosphate, 0.5 M NaCl, pH 7.5 buffer. It is aseptically filled. The protein concentration is 300-700 μg/ml (by the BCA method). For exact concentration see the product′s Certificate of Analysis.
