His Pure Cobalt Resin
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HisPur™ Cobalt Resin, Spin Columns, and Chromatography Cartridges
+
+
Co2 1
2
+
Ni2 3
4
5
+
Co2 6
L
1
2
+
Ni2 3
4
5
+
Co2 6
L
1
2
Ni2 3
4
5
6
L
Specific, fast and gentle purification of His-tagged proteins. The preferred method for purifying recombinant His-tagged proteins is immobilized metal affinity chromatography (IMAC). Traditionally, chelating chromatography resins are charged with either nickel or cobalt ions that coordinate with the histidine side chains in the 6xHis-tag. HisPur Cobalt Resin is a tetradentate chelating resin charged with cobalt that binds His-tagged proteins with high specificity and releases them under lower imidazole concentrations than required with nickel resins (see Figure). HisPur Cobalt Resin can be used to obtain high-purity proteins with no metal contamination.
Ordering Information
Highlights: • High purity – obtain pure protein without optimizing imidazole washing conditions • Specificity – cobalt:chelate binding core binds fewer contaminants, resulting in lower background than nickel (see Table) • Low metal leaching – no metal contamination in eluted sample • Versatility – purify proteins under native or denaturing conditions; compatible with cell lysis reagents and a variety of buffer additives • Flexibility – available as bulk resin or predispensed columns compatible with both spin and gravity-flow formats • Cost effective – reuse or discard • Superior – performs better than other commercially available IMAC Resins (see Figure)
kDa
M
F
1
2
Elution 3
1
2
3
4
5
Protein L
Thermo Scientific HisPur Cobalt Resin outperforms other IMAC resins. E. coli lysates containing overexpressed His-tagged green fluorescent protein (GFP), β-galactosidase or Protein L were applied to 200 μl bed volumes of HisPur Cobalt Resin and several competing cobalt and nickel resins. The first elution fraction for each IMAC resin was analyzed by SDS-PAGE and protein purity determined. Gel lanes were normalized to equivalent volume. Lane 1 = HisPur Cobalt Resin, Lane 2 = supplier C’s cobalt resin, Lane 3 = supplier S’s cobalt resin, Lane 4 = supplier G’s nickel resin, Lane 5 = supplier Q’s nickel resin, Lane 6 = Ni2+-IDA resin and Lane L = lysate load.
The HisPur Chromatography Cartridges are convenient, reliable and ready-to-use pre-packed devices for the isolation of proteins in solution and purification of His-tagged fusion proteins. The HisPur Cartridges are compatible with automated LC instrumentation, such as the ÄKTÄ™ and BioLogic Systems, and adapt to manual syringe processing.
Wash
β-gal
GFP
Product # Description
Pkg. Size
89967
HisPur Cobalt Spin Columns, 0.2 ml
25 columns
89968
HisPur Cobalt Spin Columns, 1 ml
5 columns
89969
HisPur Cobalt Spin Columns, 3 ml
5 columns
89964
HisPur Cobalt Resin
10 ml bottle
89965
HisPur Cobalt Resin
100 ml bottle
89966
HisPur Cobalt Resin
500 ml bottle
90090
HisPur Purification Kit, 0.2 ml
25 columns
90091
HisPur Purification Kit, 1 ml
5 columns
90092
HisPur Purification Kit, 3 ml
5 columns
90093
HisPur Chromatography Cartridges
5 x 1 ml
90094
HisPur Chromatography Cartridges
2 x 5 ml
L
210 110 47 32 25
6xHis-GFP
16.5
Thermo Scientific HisPur Cobalt Resin is specific for His-tagged proteins and allows mild, efficient elutions. Bacterial lysate (1.0 mg total protein) was applied to a 200 μl bed volume of HisPur Cobalt Resin in a spin column. Gel lanes were normalized to equivalent volume. M = Molecular Weight Markers (Product # 26691), L = lysate load and F = flow-through.
Thermo Scientific HisPur Cobalt Resin yields more His-tagged protein and higher purity than other Co2+ and Ni2+ IMAC Resins. His-GFP
Thermo Scientific HisPur Cobalt Resin
His-Protein L
His-β-Gal
Yield (μg)*
Purity (%)**
Yield (μg)*
Purity (%)**
Yield (μg)*
Purity (%)**
298
87
78
93
42
77
Supplier C Cobalt Resin
206
78
26
90
35
76
Supplier S Cobalt Resin
211
85
27
65
38
77
Supplier G Nickel Resin
239
84
42
83
29
68
Supplier Q Nickel Resin
242
85
24
48
30
72
Ni2+-IDA Resin
70
37
6
16
17
46
* Recovered from a 5 mg total protein load (total protein yield x purity). ** Purity of the first elution fraction.
To order, call 800-874-3723 or 815-968-0747. Outside the United States, contact your local branch office or distributor.
73
+
+
Co2 1
2
+
Ni2 3
4
5
+
Co2 6
L
1
2
+
Ni2 3
4
5
+
Co2 6
L
1
2
Ni2 3
4
5
6
L
Specific, fast and gentle purification of His-tagged proteins. The preferred method for purifying recombinant His-tagged proteins is immobilized metal affinity chromatography (IMAC). Traditionally, chelating chromatography resins are charged with either nickel or cobalt ions that coordinate with the histidine side chains in the 6xHis-tag. HisPur Cobalt Resin is a tetradentate chelating resin charged with cobalt that binds His-tagged proteins with high specificity and releases them under lower imidazole concentrations than required with nickel resins (see Figure). HisPur Cobalt Resin can be used to obtain high-purity proteins with no metal contamination.
Ordering Information
Highlights: • High purity – obtain pure protein without optimizing imidazole washing conditions • Specificity – cobalt:chelate binding core binds fewer contaminants, resulting in lower background than nickel (see Table) • Low metal leaching – no metal contamination in eluted sample • Versatility – purify proteins under native or denaturing conditions; compatible with cell lysis reagents and a variety of buffer additives • Flexibility – available as bulk resin or predispensed columns compatible with both spin and gravity-flow formats • Cost effective – reuse or discard • Superior – performs better than other commercially available IMAC Resins (see Figure)
kDa
M
F
1
2
Elution 3
1
2
3
4
5
Protein L
Thermo Scientific HisPur Cobalt Resin outperforms other IMAC resins. E. coli lysates containing overexpressed His-tagged green fluorescent protein (GFP), β-galactosidase or Protein L were applied to 200 μl bed volumes of HisPur Cobalt Resin and several competing cobalt and nickel resins. The first elution fraction for each IMAC resin was analyzed by SDS-PAGE and protein purity determined. Gel lanes were normalized to equivalent volume. Lane 1 = HisPur Cobalt Resin, Lane 2 = supplier C’s cobalt resin, Lane 3 = supplier S’s cobalt resin, Lane 4 = supplier G’s nickel resin, Lane 5 = supplier Q’s nickel resin, Lane 6 = Ni2+-IDA resin and Lane L = lysate load.
The HisPur Chromatography Cartridges are convenient, reliable and ready-to-use pre-packed devices for the isolation of proteins in solution and purification of His-tagged fusion proteins. The HisPur Cartridges are compatible with automated LC instrumentation, such as the ÄKTÄ™ and BioLogic Systems, and adapt to manual syringe processing.
Wash
β-gal
GFP
Product # Description
Pkg. Size
89967
HisPur Cobalt Spin Columns, 0.2 ml
25 columns
89968
HisPur Cobalt Spin Columns, 1 ml
5 columns
89969
HisPur Cobalt Spin Columns, 3 ml
5 columns
89964
HisPur Cobalt Resin
10 ml bottle
89965
HisPur Cobalt Resin
100 ml bottle
89966
HisPur Cobalt Resin
500 ml bottle
90090
HisPur Purification Kit, 0.2 ml
25 columns
90091
HisPur Purification Kit, 1 ml
5 columns
90092
HisPur Purification Kit, 3 ml
5 columns
90093
HisPur Chromatography Cartridges
5 x 1 ml
90094
HisPur Chromatography Cartridges
2 x 5 ml
L
210 110 47 32 25
6xHis-GFP
16.5
Thermo Scientific HisPur Cobalt Resin is specific for His-tagged proteins and allows mild, efficient elutions. Bacterial lysate (1.0 mg total protein) was applied to a 200 μl bed volume of HisPur Cobalt Resin in a spin column. Gel lanes were normalized to equivalent volume. M = Molecular Weight Markers (Product # 26691), L = lysate load and F = flow-through.
Thermo Scientific HisPur Cobalt Resin yields more His-tagged protein and higher purity than other Co2+ and Ni2+ IMAC Resins. His-GFP
Thermo Scientific HisPur Cobalt Resin
His-Protein L
His-β-Gal
Yield (μg)*
Purity (%)**
Yield (μg)*
Purity (%)**
Yield (μg)*
Purity (%)**
298
87
78
93
42
77
Supplier C Cobalt Resin
206
78
26
90
35
76
Supplier S Cobalt Resin
211
85
27
65
38
77
Supplier G Nickel Resin
239
84
42
83
29
68
Supplier Q Nickel Resin
242
85
24
48
30
72
Ni2+-IDA Resin
70
37
6
16
17
46
* Recovered from a 5 mg total protein load (total protein yield x purity). ** Purity of the first elution fraction.
To order, call 800-874-3723 or 815-968-0747. Outside the United States, contact your local branch office or distributor.
73
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