TALON Metal Affinity Resin
TALON Metal Affinity Resin
Cobalt-charged resins for high specificity purification of his-tagged proteins
Batch or gravity-flow purification of his-tagged proteins—TALON Metal Affinity Resin
TALON his-tag purification resin lets you prepare exceptionally pure his-tagged proteins from bacterial, mammalian, yeast, and baculovirus-infected cells, under native or denaturing conditions. TALON is an immobilized metal affinity chromatography (IMAC) resin charged with cobalt, which binds to his-tagged proteins with higher specificity than nickel-charged resins. As a result, TALON resin delivers his-tagged proteins of the highest purity. In addition, each cobalt ion is bound to the resin at four sites, resulting in low metal-ion leakage.
Overview
- Exhibits high affinity for his-tagged proteins
- No copurification of proteins
- Resists metal leakage
- Performs well under a wide range of purification conditions
Applications
Purified recombinant his-tagged proteins can be used for:
- Crystallography
- Functional assays
- Structural investigations
- Other applications
SDS-PAGE of TALON CellThru Resin purified proteins
SDS-PAGE of TALON CellThru Resin purified proteins. E. coli BL21 cells were sonicated in TALON wash buffer and run through a TALON CellThru column eluted in 150 mM imidazole. Note that some target protein is trapped in membrane fractions and does not get absorbed on the column. M: molecular weight marker.
His-Tag purification Columns- TALON Cobalt Resin Disposable Gravity Columns are available in 2 ml size
His-Tag purification Columns- TALON Cobalt Resin Disposable Gravity Columns are available in 2 ml size.
Native vs. denaturing purification procedures using TALON resin
Native vs. denaturing purification procedures using TALON resin.
Native purification with TALON resin preserves biological activity of proteins
Native purification with TALON resin preserves biological activity of proteins. Fresh cells (0.5 g) expressing 6xHis-GFPuv were extracted in 5 ml of 50 mM sodium phosphate; 0.3 M NaCl, pH 7.0 Panel A. Elution profile of GFP which was loaded, washed with the same buffer, and eluted with a step gradient of imidazole (150 mM). Panel B. Fractions were analyzed by SDS-PAGE. The fluorescent signal of green fluorescent protein (GFPuv) was completely enriched by TALON Superflow Resin.
Purification of 6xHis-GFPuv under denaturing conditions using TALON resin
Purification of 6xHis-GFPuv under denaturing conditions using TALON resin. The fusion protein was purified in 8 M urea using TALON resin. M=molecular weight markers.
Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol
Native purification of 6xHis protein using TALON resin in the presence of beta-mercaptoethanol. N-terminal 6xHis-tagged mouse DHFR (19.5 kDa) was expressed in E. coli. 2 ml of lysate was purified using gravity flow on TALON resin in increasing concentrations of beta-mercaptoethanol. Even lanes: 20 μl of nonadsorbed material. Odd lanes: 5 μl of eluate
Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin
Protein purification yields in the presence of beta-mercaptoethanol with TALON resin compared to Ni-NTA resin. N-terminal 6xHis DHFR was expressed and purified under native conditions. Protein concentrations were determined by Bradford assay. Yields are expressed as a percentage of total protein in the cell lysate.