HIS Lite™ Cy3 Tris NTA-Ni Complex

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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	2142.71
Solvent	Water

Spectral properties

Correction Factor (260 nm)	0.07
Correction Factor (280 nm)	0.073
Extinction coefficient (cm ^-1 M ^-1)	150000¹
Excitation (nm)	555
Emission (nm)	569
Quantum yield	0.15¹

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	Green laser
Emission	602/50 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™ Cy3 Tris NTA-Ni Complex Stock Solution

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

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

PREPARATION OF WORKING SOLUTION

HIS Lite™ Cy3 Tris NTA-Ni Complex Working Solution

Prepare a 1 to 10 µM HIS Lite™ Cy3 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™ Cy3 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™ Cy3 Tris NTA-Ni Complex working solution at the appropriate concentrations.

Note: Optimization of the HIS Lite™ Cy3 Tris NTA-Ni Complex to the His-tagged protein mix must be performed for better labeling.

Note: 1 to 10 µM of HIS Lite™ Cy3 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™ Cy3 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	46.67 µL	233.349 µL	466.699 µL	2.333 mL	4.667 mL
5 mM	9.334 µL	46.67 µL	93.34 µL	466.699 µL	933.397 µL
10 mM	4.667 µL	23.335 µL	46.67 µL	233.349 µL	466.699 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

Correction Factor (260 nm)	0.07
Correction Factor (280 nm)	0.073
Extinction coefficient (cm ^-1 M ^-1)	150000¹
Excitation (nm)	555
Emission (nm)	569
Quantum yield	0.15¹

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™ Cy3 Bis NTA-Ni Complex	555	569	150000¹	0.15¹	0.07	0.073
HIS Lite™ OG488-Tris NTA-Ni Complex	498	526	76000	-	0.31	0.12
HIS Lite™ Cy5 Tris NTA-Ni Complex	651	670	250000¹	0.27¹, 0.4²	0.02	0.03

Images

Figure 1. Cy3-Tris NTA compound is used as a sensitive fluorescent probe for detecting polyhistidine-labeled proteins in cells, solution and solid surfaces. It provides an efficient method for site-specific and stable noncovalent fluorescence labeling of polyhistidine-tagged proteins.

References

View all 50 references: Citation Explorer

Optimized immobilization of single chain variable fragment antibody onto non-toxic fluorescent nanoparticles for efficient preparation of a bioprobe.
Authors: Tateo, Seigo and Shinchi, Hiroyuki and Matsumoto, Hikaru and Nagata, Nonoka and Hashimoto, Masahito and Wakao, Masahiro and Suda, Yasuo
Journal: Colloids and surfaces. B, Biointerfaces (2023): 113192

Deep learning-assisted smartphone-based portable and visual ratiometric fluorescence device integrated intelligent gel label for agro-food freshness detection.
Authors: Lu, Zhiwei and Li, Mengjiao and Chen, Maoting and Wang, Qirui and Wu, Chun and Sun, Mengmeng and Su, Gehong and Wang, Xianxiang and Wang, Yanying and Zhou, Xinguang and Ye, Jianshan and Liu, Tao and Rao, Hanbing
Journal: Food chemistry (2023): 135640

Transition Metals Induce Quenching of Monomeric Near-Infrared Fluorescent Proteins.
Authors: Zhao, Haowen and Zastrow, Melissa L
Journal: Biochemistry (2022): 494-504

Site-Specific C-Terminal Fluorescent Labeling of Tau Protein.
Authors: Bryan, Louise and Awasthi, Saurabh and Li, Yuanjie and Nirmalraj, Peter Niraj and Balog, Sandor and Yang, Jerry and Mayer, Michael
Journal: ACS omega (2022): 47009-47014

A Simplified Protocol to Incorporate the Fluorescent Unnatural Amino Acid ANAP into Xenopus laevis Oocyte-Expressed P2X7 Receptors.
Authors: Durner, Anna and Nicke, Annette
Journal: Methods in molecular biology (Clifton, N.J.) (2022): 193-216

Construction of Fluorescent Immunosensor Quenchbody to Detect His-Tagged Recombinant Proteins Produced in Bioprocess.
Authors: Ning, Xuerao and Yasuda, Takanobu and Kitaguchi, Tetsuya and Ueda, Hiroshi
Journal: Sensors (Basel, Switzerland) (2021)

Phos-Tag Fluorescent Gel Staining for the Quantitative Detection of His- and Asp-Phosphorylated Proteins.
Authors: Kinoshita-Kikuta, Emiko and Kinoshita, Eiji and Koike, Tohru
Journal: Methods in molecular biology (Clifton, N.J.) (2021): 73-78

Affinity Purification of GO-Matryoshka Biosensors from E. coli for Quantitative Ratiometric Fluorescence Analyses.
Authors: Sadoine, Mayuri and Castro-Rodríguez, Vanessa and Poloczek, Tobias and Javot, Helene and Sunal, Erdem and Wudick, Michael M and Frommer, Wolf B
Journal: Bio-protocol (2020): e3773

Ultrafast in-gel detection by fluorescent super-chelator probes with HisQuick-PAGE.
Authors: Brüchert, Stefan and Joest, Eike F and Gatterdam, Karl and Tampé, Robert
Journal: Communications biology (2020): 138

Quantitative monitoring of His and Asp phosphorylation in a bacterial signaling system by using Phos-tag Magenta/Cyan fluorescent dyes.
Authors: Kinoshita-Kikuta, Emiko and Kusamoto, Hiroshi and Ono, Syogo and Akayama, Keisuke and Eguchi, Yoko and Igarashi, Masayuki and Okajima, Toshihide and Utsumi, Ryutaro and Kinoshita, Eiji and Koike, Tohru
Journal: Electrophoresis (2019): 3005-3013