||Recombinant Human Eukaryotic Initiation Factor 2-alpha produced inSf9is a non-glycosylated, polypeptide chain having a molecular mass of 36 kDa. EIF2-is purified by proprietary chromatographic techniques.
||The translation initiation factor eIF2 catalyzes the first regulated step of protein synthesis initiation, promoting the binding of the initiator tRNA to 40S ribosomal subunits. Binding occurs as a ternary complex of methionyl-tRNA, eIF2, and GTP. eIF2 is composed of 3 nonidentical subunits, alpha (36 kD), beta (38 kD, MIM 603908), and gamma (52 kD, MIM 300161). The rate of formation of the ternary complex is modulated by the phosphorylation state of eIF2-alpha (Ernst et al., 1987).
||Baculovirus, SF9 Insect Cells.
||Sterile Filtered White lyophilized (freeze-dried) powder.
||Greater than 95% as determined by SDS-PAGE.
||EIF2A is supplied as lyophilized freeze dry powder without additives.
||We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute lyophilized recombinant human eIF2-α in phosphate buffered saline, to a concentration of 0.2-1.0 mg/mL. Following reconstitution, the protein solution should be supplemented with 1mM β-mercaptoethanol (final concentration) and 10% glycerol (final concentration) to prevent precipitation.
||Recombinant human eIF2-α is phosphorylatable in vitro, using either immunoprecipitated active PKR or extracts from IFN-γ stimulated HEK 293 cells. This phosphorylation can be monitored by Western blot analysis using phosphorylation site specific antibody directed to eIF2-α [pS51] in conjunction with chemiluminescence detection methods. Please note: Kinase activity may vary depending on the substrate and reaction conditions.
||Store at 4°C if entire vial will be used within 1-2 weeks. Store, frozen at -20°C for longer periods of time. Avoid multiple freeze-thaw cycles.