URL: http://www.addgene.org/124296
Proper Citation: RRID:Addgene_124296
Insert Name: His6 tag - TEV protease site - T7 tag - Streptavidin (15-159) - superfolder GFP
Organism: Synthetic
Bacterial Resistance: Kanamycin
Vector Backbone Description: Backbone Marker:Novagen; Backbone Size:5237; Vector Backbone:pET29b; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: T7 promoter so expression requires DE3 strain. Pilot experiments using E. coli BL21(DE3) indicate 18C expression overnight gives higher yields than 37C (for 4 hours). The fusion protein was soluble and purified by Ni-NTA chromatography using 50 mM Tris pH 8.0, 300 mM NaCl, 10/300 mM imidazole. Size exclusion chromatography indicated that the fusion protein was tetrameric in Tris-buffered saline; no aggregates were present.
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Source: Addgene