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HisPur Cobalt Resin outperforms other IMAC resins
Comparable yield and higher purity obtained with HisPur Cobalt Resin (lane 1) compared to other IMAC resins (lanes 2 to 6). Cell lysates containing over-expressed recombinant 6xHis-tagged protein were prepared in B-PER Bacterial Protein Extraction Reagent (Part No. 78243) and protease inhibitors. Protein concentrations were determined by Coomassie Plus Protein Assay (Part No. 23238). E. coli lysates containing over-expressed His-tagged GFP, ß-galactosidase or Protein L were applied to 0.2mL bed volumes of each IMAC resin in spin column format. Binding, wash and elution buffers were prepared and used per each manufacturer's instructions. The first elution fraction for each IMAC resin was analyzed by SDS-PAGE and protein purity determined by densitometry. Gel lanes were normalized to equivalent volume. Lanes: 1= HisPur Cobalt Resin, 2= supplier C cobalt resin, 3= supplier S cobalt resin, 4= supplier G nickel resin, 5= supplier Q nickel resin, 6= Ni-IDA and L= lysate load.
