This Mouse IFNAR1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of IFNAR1 protein (Cat: 50469-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of mouse IFNAR1 (NP_034638.2) extracellular domain (Met 1-Thr 429) was expressed, with a polyhistidine tag at the C-terminus.
The recombinant mouse IFNAR1 consists of 414 amino acids and has a predicted molecular mass of 47 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rm IFNAR1 is approximately 55-60 kDa due to glycosylation.
Mouse IFNAR1 HEK293 Overexpression Lysate: 用法
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Interferon-alpha/beta receptor alpha chain (IFNAR1) is a type I membrane protein that forms one of the two chains of a receptor for interferons alpha and beta. Binding and activation of the receptor stimulates Janus protein kinases, which in turn phosphorylate several proteins, including STAT1 and STAT2. The encoded protein also functions as an antiviral factor. Tyk2 slows down IFNAR1 degradation and that this is due, at least in part, to inhibition of IFNAR1 endocytosis. Mutant versions of IFNAR1, in which Tyr466 is changed to phenylalanine, can act in a dominant negative manner to inhibit phosphorylation of STAT2. These observations are consistent with a model in which IFNAR1 mediates the interaction between JAK kinases and the STAT transcription factors.