HIS Lite™ iFluor™ 647 Tris NTA-Ni Complex

Ordering information

Price
Catalog Number
Unit Size
Quantity

Add to cart

Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	2094.58
Solvent	Water

Spectral properties

Excitation (nm)	648
Emission (nm)	668

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure

Overview SDS Protocol

Platform

Gel Imager

Excitation	Red laser
Emission	700/50 nm

Example protocol

PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

HIS Lite™ iFluor® 647 Tris NTA-Ni Complex Stock Solution

Prepare a 5 to 10 mM stock solution by adding an appropriate amount of ddH2O.

Note: Store any unused stock solution at -20 °C. Avoid repeated freeze-thaw cycles and minimize light exposure.

PREPARATION OF WORKING SOLUTION

HIS Lite™ iFluor® 647 Tris NTA-Ni Complex Working Solution

Prepare a 1 to 10 µM HIS Lite™ iFluor® 647 Tris NTA-Ni Complex working solution in PBS.

Note: Ensure that there is sufficient working solution to fully submerge the gel. After use, discard the working solution. Do not reuse.

SAMPLE EXPERIMENTAL PROTOCOL

The following protocol should be used only as a guideline and may require optimization to better suit your specific experimental needs.

Post-run Gel Staining Protocol

Run gels based on your standard protocol.
Place the gel in a suitable container. Fix the gel in the fixing solution for 60 minutes. Note: 40% ethanol + 10% acetic acid can be used as a fixing solution.
Wash the gel twice with the ultra-pure water.
Incubate the gel in the HIS Lite™ iFluor® 647 Tris NTA-Ni Complex working solution for 60 minutes.

Note: Be sure to fully submerge the gel in the working solution.
Remove the working solution and wash the gel twice with PBS.
Proceed to imaging the gel immediately.

For In Vitro Complex Formation

Mix the His-tagged protein solution and the HIS Lite™ iFluor® 647 Tris NTA-Ni Complex working solution at the appropriate concentrations.

Note: Optimization of the HIS Lite™ iFluor® 647 Tris NTA-Ni Complex to the His-tagged protein mix must be performed for better labeling.

Note: 1 to 10 µM of HIS Lite™ iFluor® 647 Tris NTA-Ni Complex can be used as a starting concentration.

Note: The reaction can be performed in a buffer containing 50 mM HEPES/KOH, pH 7.4, 100 mM KCl, 1 mM MgCl2, 2 mM β-mercaptoethanol, 5% glycerol, or a buffer of your choice.
Mix can be incubated for 30 minutes at room temperature or 4 ℃.

Note: Optimization of the incubation time and conditions must be performed for better labeling
Mix can then be subjected to column purification or any other downstream process.

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of HIS Lite™ iFluor™ 647 Tris NTA-Ni Complex to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	47.742 µL	238.711 µL	477.423 µL	2.387 mL	4.774 mL
5 mM	9.548 µL	47.742 µL	95.485 µL	477.423 µL	954.845 µL
10 mM	4.774 µL	23.871 µL	47.742 µL	238.711 µL	477.423 µL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
	/		=		x		=

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Excitation (nm)	648
Emission (nm)	668

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Quantum yield	Correction Factor (260 nm)	Correction Factor (280 nm)
HIS Lite™ iFluor® 568 Tris NTA-Ni Complex	568	587	100000¹	0.57¹	0.34	0.15

Images

Figure 1. Fluorescent tris-NTA compounds provide an efficient method for site-specific and stable noncovalent fluorescence labeling of polyhistidine-tagged proteins. iFluor 647-Tris NTA compound is used as a sensitive fluorescent probe for detecting polyhistidine-labeled proteins in cells, solution and solid surfaces. It has much stronger fluorescence than the other similar wavelength of NTA compounds.

References

View all 44 references: Citation Explorer

Selective APC-targeting of a novel Fc-fusion multi-immunodominant recombinant protein (tTax-tEnv:mFcγ2a) for HTLV-1 vaccine development.
Authors: Shafifar, Mina and Mozhgani, Sayed-Hamidreza and Razavi Pashabayg, Kobra and Mosavat, Arman and Karbalaei, Mohsen and Norouzi, Mehdi and Rezaee, Seyed Abdolrahim
Journal: Life sciences (2022): 120920

Simplified detection of polyhistidine-tagged proteins in gels and membranes using a UV-excitable dye and a multiple chelator head pair.
Authors: Raducanu, Vlad-Stefan and Isaioglou, Ioannis and Raducanu, Daniela-Violeta and Merzaban, Jasmeen S and Hamdan, Samir M
Journal: The Journal of biological chemistry (2020): 12214-12223

A bispecific circular aptamer tethering a built-in universal molecular tag for functional protein delivery.
Authors: Pan, Xiaoshu and Yang, Yu and Li, Long and Li, Xiaowei and Li, Qiang and Cui, Cheng and Wang, Bang and Kuai, Hailan and Jiang, Jianhui and Tan, Weihong
Journal: Chemical science (2020): 9648-9654

The surface syndecan protein from Macrobrachium rosenbergii could function as mediator in bacterial infections.
Authors: Yang, Hui and Xiong, Haoran and Mi, Kaihang and Zhang, Yingying and Zhang, Xiaojun and Chen, Guohong
Journal: Fish & shellfish immunology (2020): 62-68

Reshaping nanobodies for affinity purification on protein a.
Authors: Crauwels, Maxine and Van Vaerenbergh, Nele and Kulaya, Neeme Benedict and Vincke, Cécile and D'Huyvetter, Matthias and Devoogdt, Nick and Muyldermans, Serge and Xavier, Catarina
Journal: New biotechnology (2020): 20-28

[Cloning and expression of SmDXS2 gene in Swertia mussotii].
Authors: Li, Wen-Jing and Xiang, Bei-Bei and Sun, Yan-Xiang and Hou, Xiao-Qiang and Han, Mei-Ling and Li, Xiao-Xue and Wang, Yong and Guo, Shuo
Journal: Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica (2019): 935-941

Overexpression, Purification and Functional Characterisation of Wild-Type HIV-1 Subtype C Protease and Two Variants Using a Thioredoxin and His-Tag Protein Fusion System.
Authors: Zondagh, Jake and Williams, Alison and Achilonu, Ikechukwu and Dirr, Heini W and Sayed, Yasien
Journal: The protein journal (2018): 369-379

Molecular characterization of Babesia microti thioredoxin (BmTrx2) and its expression patterns induced by antiprotozoal drugs.
Authors: Huang, Jingwei and Xiong, Kang and Zhang, Houshuang and Zhao, Yanzhen and Cao, Jie and Gong, Haiyan and Zhou, Yongzhi and Zhou, Jinlin
Journal: Parasites & vectors (2018): 38

Characterization of Recombinant His-Tag Protein Immobilized onto Functionalized Gold Nanoparticles.
Authors: Torres-González, Lisa and Díaz-Ayala, Ramonita and Vega-Olivencia, Carmen A and López-Garriga, Juan
Journal: Sensors (Basel, Switzerland) (2018)

Facile fabrication of nickel immobilized on magnetic nanoparticles as an efficient affinity adsorbent for purification of his-tagged protein.
Authors: Rashid, Zahra and Naeimi, Hossein and Zarnani, Amir-Hassan and Mohammadi, Fereshteh and Ghahremanzadeh, Ramin
Journal: Materials science & engineering. C, Materials for biological applications (2017): 670-676