Fusion Protein Purification, 25217 HisPur Ni-NTA Superflow Agarose

We offer a variety of purification resins for the purification of recombinant proteins from cultures such as E. coli or Picchia. These resins are available in multiple formats to accommodate a variety of needs, from screening, batch, pilot, and process purification. Superflow agarose resins have undergone extensive chemical characterization.

  • Product choice—a variety of ligand and support choices to meet purification goals
  • High performance—resins that maximize binding capacity, purity, batch size, and/or processing time
  • Flexible—available in multiple formats, including bulk resin, spin columns and kits, FPLC cartridges, and 96-well spin plates

Choose the right resin for your recombinant protein purification needs

  HisPur Ni-NTA Agarose Resin HisPur Ni-NTA Superflow Resin HisPur Cobalt Agarose Resin HisPur Cobalt Superflow Resin Glutathione Agarose Resin Glutathione Superflow Resin
Target His-tagged fusion proteins His-tagged fusion proteins His-tagged fusion proteins His-tagged fusion proteins GST-fusion proteins GST-fusion proteins
Static binding capacity† ~60 mg/mL >60 mg/mL ≥15 mg/mL >30 mg/mL ~40 mg/mL ~30 mg/mL
Dynamic binding capacity† 18 mg/mL 20 mg/mL ND* >20 mg/mL ~10.5 mg/mL ~10 mg/mL
Purity High High Higher Higher High High
Maximum flow rates 800 cm/hr 1,200 cm/hr 800 cm/hr 1,200 cm/hr 800 cm/hr 1,200 cm/hr
Maximum pressure 0.35 MPa 0.65 MPa 0.35 MPa 0.65 MPa 0.35 MPa 0.65 MPa
Support 6% agarose 6% agarose, highly crosslinked 6% agarose 6% agarose, highly crosslinked 6% agarose 6% agarose, highly crosslinked
Application scales Screening, batch, pilot Batch, pilot, process Screening, batch, pilot Batch, pilot, process Screening, batch, pilot Batch, pilot, process
No. of reuses 5 25 5 25 5 25
  88221 (10 mL),
88222 (100 mL), 88223 (500 mL)
25214 (10 mL), 25215 (50 mL), 25216 (250 mL), 25217 (1 L) 89964 (10 mL), 89965 (100 mL), 89966 (500 mL) 25228 (10 mL), 25229 (50 mL), 25230 (250 mL), 25231 (1 L) 16100 (10 mL), 16101 (100 mL), 16102 (500 mL) 25236 (10 mL), 25237 (50 mL), 25238 (250 mL), 25239 (1 L)
†See product pages for details.
*Not determined.

Smaller-scale purification solutions

In addition to bulk agarose-based resins, we offer convenient agarose-based spin columns (see table below) for rapid sample processing.  The spin columns are also available in a kit format that includes all buffers needed for binding and elution. The FPLC columns are compatible with the most popular automated FPLC systems. The resins are also supplied in convenient 96-well filter plates for high-throughput sample processing. 

Choose the small-scale purification product that fits your experiment

Resin Format Resin volume (mL) Catalog #
Ni-NTA Spin Column 0.2 88224
Ni-NTA Spin Column 1 88225
Ni-NTA Spin Column 3 88226
Ni-NTA Spin Column Kit 0.2 88227
Ni-NTA Spin Column Kit 1 88228
Ni-NTA Spin Column Kit 3 88229
Ni-NTA Chromatography Cartridge 1 90098
Ni-NTA Chromatography Cartridge 5 90099
Ni-NTA 96-well Spin Plate 0.1 88230
Cobalt Spin Column 0.2 89967
Cobalt Spin Column 1 89968
Cobalt Spin Column 3 89969
Cobalt Spin Column Kit 0.2 90090
Cobalt Spin Column Kit 1 90091
Cobalt Spin Column Kit 3 90092
Cobalt Chromatography Cartridge 1 90093
Cobalt Chromatography Cartridge 5 90094
Cobalt 96-well Spin Plate 0.1 90095
Glutathione Spin Column 0.2 16103
Glutathione Spin Column 1 16104
Glutathione Spin Column 3 16105
Glutathione Spin Column Kit 0.2 16106
Glutathione Spin Column Kit 1 16107
Glutathione Spin Column Kit 3 16108
Glutathione Chromatography Cartridge 1 16109
Glutathione Chromatography Cartridge 5 16110
Glutathione 96-well Spin Plate 0.1 16111
High-yield, high-purity, medium-scale purification of 6xHis-Tagged protein with HisPur Ni-NTA Superflow Agarose
High-yield, high-purity, medium-scale purification of 6xHis-Tagged protein. More than 4 grams of over-expressed 6xHis-GFP were purified in 3 hours using 200 mL columns containing HisPur Ni-NTA Superflow Agarose or Qiagen™ Ni-NTA Superflow. One liter of lysate was loaded at a flow rate of 20 mL/min, then washed until baseline with wash buffer containing 30 mM imidazole. Bound protein was eluted with buffer containing 300 mM imidazole. Fractions containing purified 6xHis-GFP were pooled and quantitated using Pierce 660 nm Protein Assay (Cat. No. 22662). Load, flow-through, wash, and eluate fractions were separated by SDS-PAGE, stained with Imperial Protein Stain (Cat. No. 24615) and evaluated using myImageAnalysis Software (Cat. No. 62237) to determine purity.

Thermo Scientific Pierce Glutathione Agarose delivers high yield and high purity GST-fusion proteins. E. coli lysate (14.4 mg total protein) containing overexpressed GST was incubated with 50 µL GSH resin from various suppliers and purified per manufacturers' instructions. The amount of GST eluted from the resin (yield) was quantified by Thermo Scientific™ Coomassie Plus Protein Assay. Purity was assessed by densitometry of the stained gel lanes. M= MW marker; L=Lysate load; FT=Flow-through; E=Elution.

Other recombinant protein purification resins

Other popular tags for mammalian system protein expression include HA or c-Myc. These tags are extremely useful for protein enrichment applications, such as immunoprecipitation (IP) or co-immunoprecipitation (co-IP).

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