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HIS Lite™ iFluor™ 647 Tris NTA Chelator

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Physical properties
Molecular weight1699.84
SolventDMSO
Spectral properties
Correction Factor (260 nm)0.03
Correction Factor (280 nm)0.03
Correction Factor (656 nm)0.0793
Extinction coefficient (cm -1 M -1)2500001
Excitation (nm)656
Emission (nm)670
Quantum yield0.251
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
1699.84
Correction Factor (260 nm)
0.03
Correction Factor (280 nm)
0.03
Correction Factor (656 nm)
0.0793
Extinction coefficient (cm -1 M -1)
2500001
Excitation (nm)
656
Emission (nm)
670
Quantum yield
0.251
iFluor® 647 Tris-NTA Chelator is used as a sensitive red fluorescent probe for detecting polyhistidine-labeled proteins in combination with the addition of a heavy metal ion such as Ni2+ in cells, solution and solid surfaces. Fluorescent tris-NTA compounds provide an efficient method for site-specific and stable noncovalent fluorescence labeling of polyhistidine-tagged proteins. In contrast to the transient binding of conventional mono-NTA, the multivalent interaction of tris-NTA conjugated fluorophores form a much more stable complex with polyhistidine-tagged proteins. The high selectivity of tris-NTA compounds toward cumulated histidines enables the selective labeling of proteins in cell lysates and on the surface of live cells. Fluorescent tris-NTA conjugates can be applied for the analysis of a ternary protein complex in solution and on surfaces. In combination with OG488-tris-NTA compound iFluor® 647 Tris-NTA can be used for multicolor analysis of polyhistidine-tagged proteins. AAT Bioquest also offers HIS Lite™ iFluor™ 647 Tris NTA-Ni Complex that can be directly used for the specific and highly sensitive detection of His-tagged fusion proteins without the addition of a heavy metal ion. The transition metal ions (e.g., Ni ion)-mediated complexation of polyhistidine-labeled proteins with fluorescent tris-NTA conjugates provides a sensitive reporter for detecting and monitoring protein-protein interactions in real time.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of HIS Lite™ iFluor™ 647 Tris NTA Chelator 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 mM58.829 µL294.145 µL588.291 µL2.941 mL5.883 mL
5 mM11.766 µL58.829 µL117.658 µL588.291 µL1.177 mL
10 mM5.883 µL29.415 µL58.829 µL294.145 µL588.291 µL

Molarity calculator

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Spectrum


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spectrum

Spectral properties

Correction Factor (260 nm)0.03
Correction Factor (280 nm)0.03
Correction Factor (656 nm)0.0793
Extinction coefficient (cm -1 M -1)2500001
Excitation (nm)656
Emission (nm)670
Quantum yield0.251

Product Family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)
HIS Lite™ iFluor™ 568 Tris NTA Chelator56858710000010.5710.340.15

Images


References


View all 50 references: Citation Explorer
Correction: Diamond nanowires modified with poly[3-(pyrrolyl)carboxylic acid] for the immobilization of histidine-tagged peptides.
Authors: Subramanian, Palaniappan and Mazurenko, Ievgen and Zaitsev, Vladimir and Coffinier, Yannick and Boukherroub, Rabah and Szunerits, Sabine
Journal: The Analyst (2024)
Analysis of histidine-tagged recombinant proteins from nickel and copper coated surfaces by direct electrospray ionization and desorption electrospray ionization mass spectrometry.
Authors: Javanshad, Roshan and Taylor, Christopher James and Delavari, Niusha and Barkman, Todd J and Stull, Frederick and Venter, Andre R
Journal: Rapid communications in mass spectrometry : RCM (2023): e9516
Poly-Histidine-Tagged Protein Purification Using Immobilized Metal Affinity Chromatography (IMAC).
Authors: Loughran, Sinéad T and Bree, Ronan T and Walls, Dermot
Journal: Methods in molecular biology (Clifton, N.J.) (2023): 193-223
Magnetically purification/immobilization of poly histidine-tagged proteins by PEGylated magnetic graphene oxide nanocomposites.
Authors: Khataminezhad, Ehteram Sadat and Hajihassan, Zahra and Razi Astaraei, Fatemeh
Journal: Protein expression and purification (2023): 106264
Chromatographic purification of histidine-tagged proteins using zirconia particles modified with phosphate groups.
Authors: Kanoh, Shogo and Shiraki, Kentaro and Wada, Momoyo and Tanaka, Takeshi and Kitamura, Masahiro and Kato, Katsuya and Hirano, Atsushi
Journal: Journal of chromatography. A (2023): 464112
Synthesis of micron-sized magnetic agarose beads chelated with nickel ions towards the affinity-based separation of histidine-tagged/rich proteins.
Authors: Zhao, Ya-Qi and Yu, Shi-Song and Chen, Meng-Ying and Wang, Yuan and Shi, Yu-Jun and Wang, Xin-Yu and Zhao, Jia-Meng and Dong, Lin-Yi and Zhao, Zhen-Yu and Wang, Xian-Hua
Journal: Journal of chromatography. A (2023): 464365
Cation affinity purification of histidine-tagged proteins.
Authors: Sun, Hongxu and Wang, Hongrui and Chen, Qiwei and Dong, Wenge and Gao, Chao and Song, Haiyan and Peng, Hui and Li, Ren and Wu, Hao and Hou, Liangyu and Chang, Yanhong and Luo, Hui
Journal: Applied microbiology and biotechnology (2023): 2639-2651
Synthesis of Ni2+-functionalized polydopamine magnetic beads for facilitated purification of histidine-tagged proteins.
Authors: Shariati, Alireza and Hosseinzadeh, Sara Ali and Barghi, Zahra and Mortazavi, Sogand Sadat and Atarod, Kosar and Shariati, Fatemeh Sadat and Farahmand, Behrokh
Journal: AMB Express (2023): 112
Automated ssDNA SELEX Using Histidine-Tagged Target Proteins.
Authors: Breuers, Stefan and Mayer, Günter
Journal: Methods in molecular biology (Clifton, N.J.) (2023): 3-11
Ni(II) functionalized polyhedral oligomeric silsesquioxane based capillary monolith for purification of histidine-tagged proteins by immobilized metal affinity micro-chromatography.
Authors: Çambay Kuban, Fatma and Koçer, İlkay and Kip, Çiğdem and Çelik, Eda and Tuncel, Ali
Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2023): 123759