biotechrabbit™ Tris-NTA Biotin for high-affinity His-tag binding.
His-tags are one of the most commonly used tags for protein expression analysis. Conventional metal ion chelators, such as nitrilotriacetic acid (NTA) and iminodiacetic acid (IDA), bind His-tags with low affinities in the range of 10 µM. The biotechrabbit Tris-NTA complexes three NTA groups that together bind a 6×His-tag with an affinity that is four orders of magnitude higher (1 nM) than is possible with conventional chelators (10 µM). The binding of His-tags is stoichiometric and single-molecule detection is possible. Binding is reversible: bound His-tags can be released with imidazole or ethylenediaminetetraacetic acid (EDTA).
biotechrabbit™ Tris-NTA is available with a free amine group or conjugated to biotin. It can be used in a large range of applications, including protein detection and labeling, coupling proteins, lipids or cells to surfaces, protein purification and reversible protein modification.
NOTICE TO PURCHASER: LIMITED LICENSE
The use of this product for research purposes is covered by a license from QIAGEN Group. No rights to use this product to perform or offer diagnostic, companion diagnostic, commercial testing or other commercial services for money or money’s worth are granted by the supply of this product expressly, by implication or by estoppel.
Should you wish to use this product for any other purpose not covered by this license, please contact QIAGEN Corporate Business Development at bd@qiagen.com.
Component | Composition |
Tris-NTA Biotin | Tris-NTA Biotin (1 mg/ml) in PBS |
STORAGE | 4°C (until expiry date – see product label) |
CHEMICAL NAME | Biotin-Tris-NTA |
STABILITY | 24 months |
STORAGE CONDITIONS | store at 4°C |
METHOD FOR DETERMINING IDENTITY | HPLC, Mass spec |
CAS NUMBER | free base: 1070867-85-4 |
MOLUCULAR FORMULAR | free base: C59H93N11O25S |
MOLECULAR WEIGHT | free base: 1388.49 |
SOURCE | Synthetic |
PURITY | >95% (HPLC) |
FORM | 1 mg/ml PBS solution |
Quality Control
Identity
Identity of the substance was determined by MS analysis. The identity was consistent with the reference substance.
Purity
The purity of Tris-NTA Biotin was determined by HPLC. Purity was >95%.
Tris-NTA is not pre-loaded with a metal ion.
Please, choose the metal ion (i.e. Nickel or Cobalt) and loading method that fit best to your application.
As an example, please, refer to Figure 5 in "Reichel et al., Anal Chem., 2007, 79, 8590–600":
Ni2+ loading by injection of 10 mM NiCl2 when using Tris-NTA with a biosensor surface.
Application examples
High-Affinity Adaptors for Switchable Recognition of Histidine-Tagged Proteins.
Lata et al., J. Am. Chem. Soc., 2005, 127, 10205–10215
Specific and Stable Fluorescence Labeling of Histidine-Tagged Proteins for Dissecting Multi-Protein Complex Formation.
Lata et. al., J. Am. Chem. Soc., 2006, 128, 2365–2372
Noncovalent, Site-Specific Biotinylation ofHistidine-Tagged Proteins.
Reichel et al., Anal Chem., 2007, 79, 8590–600
Identifying Modulators of Protein-Protein Interactions Using Photonic Crystal Biosensors.
Heeres et al., J Am Chem Soc. 2009, 131: 18202–18203
Tris-Nitrilotriacetic Acids of Sub-nanomolar Affinity Toward Hexahistidine Tagged Molecules.
Huang et al., Bioconjug Chem., 2009, 20: 1667–1672
Four-color single-molecule fluorescence with noncovalent dye labeling to monitor dynamic multimolecular complexes.
DeRocco et al., BioTechniques 2010, 49: 807-816
Co- and distinct existence of Tris-NTA and biotin functionalities on individual and adjacent micropatterned surfaces generated by photo-destruction.
Biswas et al., Soft Matter, 2014, 10, 2341–2345
High-affinity gold nanoparticle pin to label and localize histidine tagged protein in macromolecular assemblies.
Anthony et al., Structure, 2014, 22: 628–635
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