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6XHis Succinimidyl Ester

Purification test of 6xHis-tagged IgG with His SpinTrap™. IgG was labeled with 6xHis,SE and then loaded onto His SpinTrap™. The experiment showed that 547.6 μg of IgG-His could bind to His SpinTrap™, and 457.7 μg was recovered (84%) using 250 mM Imidazole Elution Buffer.
Purification test of 6xHis-tagged IgG with His SpinTrap™. IgG was labeled with 6xHis,SE and then loaded onto His SpinTrap™. The experiment showed that 547.6 μg of IgG-His could bind to His SpinTrap™, and 457.7 μg was recovered (84%) using 250 mM Imidazole Elution Buffer.
Purification test of 6xHis-tagged IgG with His SpinTrap™. IgG was labeled with 6xHis,SE and then loaded onto His SpinTrap™. The experiment showed that 547.6 μg of IgG-His could bind to His SpinTrap™, and 457.7 μg was recovered (84%) using 250 mM Imidazole Elution Buffer.
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Physical properties
Molecular weight1495.55
SolventDMSO
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure

OverviewpdfSDSpdfProtocol


Molecular weight
1495.55
6XHis Succinimidyl Ester is an excellent building block to make 6XHis conjugates for developing His tag detection probes and purification tools. It readily reacts with a biomolecule that contains an amino group such as antibodies. The 6xHis tag is the most common his-tag and has a molecular weight of ~0.8kDa. His-tags (i.e., polyhistidine tags) comprise a consecutive series of six to ten histidine residues. His tags are used for many recombinant proteins to facilitate purification, allowing researchers to extract a protein of interest from thousands of other proteins found in a cell or cell lysate. The small size of the 6x-His tag has a lower possibility for the tag to affect the functionality of the fusion protein. Histidine forms coordination bonds with immobilized transition metal ions, and this property can be utilized for protein purification. His-tag protein purification is by a specialized form of affinity chromatography, called immobilized metal affinity chromatography (IMAC), where proteins or peptides are separated according to their affinity for metal ions immobilized to a solid chelating resin. During this process, a small His-tag is fused to either the N or C terminus of the target protein, enabling capture by nickel or cobalt ions coordinated on a variety of resins. The small size, low cost, and ease of use have made the His-tag the most popular affinity-tag available. AAT Bioquest offers the largest collection of 6XHis, NTA, bis-NTA, tris-NTA and IDA reagents for His Tag detections and purifications.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of 6XHis Succinimidyl Ester to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM66.865 µL334.325 µL668.65 µL3.343 mL6.687 mL
5 mM13.373 µL66.865 µL133.73 µL668.65 µL1.337 mL
10 mM6.687 µL33.433 µL66.865 µL334.325 µL668.65 µL

Molarity calculator

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References


View all 19 references: Citation Explorer
Selective Acylation of Proteins at Gly and Lys in His Tags.
Authors: Jensen, Knud J and Thygesen, Mikkel B and Sørensen, Kasper K and Wu, Shunliang and Treiberg, Tuule and Schoffelen, Sanne
Journal: Chembiochem : a European journal of chemical biology (2022): e202200359
Poly-Gly Region Regulates the Accessibility of Metal Binding Sites in Snake Venom Peptides.
Authors: Wa Tły, Joanna and Hecel, Aleksandra and Wieczorek, Robert and Rowińska-Żyrek, Magdalena and Kozłowski, Henryk
Journal: Inorganic chemistry (2022): 14247-14251
Engineering Polyhistidine Tags on Surface Proteins of Acidithiobacillus ferrooxidans: Impact of Localization on the Binding and Recovery of Divalent Metal Cations.
Authors: Jung, Heejung and Inaba, Yuta and Jiang, Virginia and West, Alan C and Banta, Scott
Journal: ACS applied materials & interfaces (2022): 10125-10133
Recombinant overexpression of camel hepcidin cDNA in Pichia pastoris: purification and characterization of the polyHis-tagged peptide HepcD-His.
Authors: Boumaiza, Mohamed and Chahed, Haifa and Ezzine, Aymen and Jaouen, Maryse and Gianoncelli, Alessandra and Longhi, Giovanna and Carmona, Fernando and Arosio, Paolo and Sari, Marie-Agnès and Marzouki, Mohamed Nejib
Journal: Journal of molecular recognition : JMR (2018): e2729
Recombinant overexpression of camel hepcidin cDNA in Pichia pastoris: purification and characterization of the polyHis-tagged peptide HepcD-His.
Authors: Boumaiza, Mohamed and Chahed, Haifa and Ezzine, Aymen and Jaouen, Maryse and Gianoncelli, Alessandra and Longhi, Giovanna and Carmona, Fernando and Arosio, Paolo and Sari, Marie-Agnès and Marzouki, Mohamed Nejib
Journal: Journal of molecular recognition : JMR (2017)
The unusual metal ion binding ability of histidyl tags and their mutated derivatives.
Authors: Brasili, Davide and Watly, Joanna and Simonovsky, Eyal and Guerrini, Remo and Barbosa, Nuno A and Wieczorek, Robert and Remelli, Maurizio and Kozlowski, Henryk and Miller, Yifat
Journal: Dalton transactions (Cambridge, England : 2003) (2016): 5629-39
African Viper Poly-His Tag Peptide Fragment Efficiently Binds Metal Ions and Is Folded into an α-Helical Structure.
Authors: Watly, Joanna and Simonovsky, Eyal and Barbosa, Nuno and Spodzieja, Marta and Wieczorek, Robert and Rodziewicz-Motowidlo, Sylwia and Miller, Yifat and Kozlowski, Henryk
Journal: Inorganic chemistry (2015): 7692-702
His-tag binding by antibody C706 mimics β-amyloid recognition.
Authors: Teplyakov, Alexey and Obmolova, Galina and Canziani, Gabriela and Zhao, Yonghong and Gutshall, Lester and Jung, Sonia S and Gilliland, Gary L
Journal: Journal of molecular recognition : JMR (2011): 570-5
The venom of the snake genus Atheris contains a new class of peptides with clusters of histidine and glycine residues.
Authors: Favreau, Philippe and Cheneval, Olivier and Menin, Laure and Michalet, Sophie and Gaertner, Hubert and Principaud, Franck and Thai, Robert and Ménez, André and Bulet, Philippe and Stöcklin, Reto
Journal: Rapid communications in mass spectrometry : RCM (2007): 406-12
Cleavage of recombinant proteins at poly-His sequences by Co(II) and Cu(II).
Authors: Andberg, Martina and Jäntti, Jussi and Heilimo, Sara and Pihkala, Päivi and Paananen, Arja and Koskinen, Ari M P and Söderlund, Hans and Linder, Markus B
Journal: Protein science : a publication of the Protein Society (2007): 1751-61