HIS Lite™ Cy5 Bis NTA-Ni Complex

Polyhistidine is one of the most popular affinity tags incorporated into recombinant proteins. It can be inserted either at the N- or C-terminus, and expressed in a variety of hosts. Due to its small size, the polyhistidine tag serves as an elegant tool for both protein purification and detection. HIS Lite™ Cy3 Bis NTA-Ni and Cy5 Bis NTA-Ni Complexes provide specific and highly sensitive detection of His-tagged fusion proteins. The Ni-NTA complexes were first reported by Kapanidis et Al. to be specific for polyhistidine tags with minimal crossreactivity. Cy3 and Cy5 dyes demonstrate strong fluorescent signals at commonly available wavelengths and with little quenching. The Cy3 Bis NTA-Ni and Cy5 Bis NTA-Ni Complexes can be directly applied either to an SDS-PAGE gel or Western blot membrane for fluorescence imaging. Detection with the Cy3 Bis NTA-Ni and Cy5 Bis NTA-Ni Complexes requires less incubation time than for protein-antibody binding. No secondary reaction is required since the Ni-NTA complexes are directly conjugated to the fluorophores.

Protocol

SDS

COA

Catalog	Size	Price	Quantity
12613	1 mg	Price

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of HIS Lite™ Cy5 Bis 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	66.665 µL	333.327 µL	666.653 µL	3.333 mL	6.667 mL
5 mM	13.333 µL	66.665 µL	133.331 µL	666.653 µL	1.333 mL
10 mM	6.667 µL	33.333 µL	66.665 µL	333.327 µL	666.653 µL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	1500.03 g/mol	=	mL	x	mM	=	mmol

Citations

View all 2 citations: Citation Explorer

ACE2 Receptor and Antibody Binding to SARS-CoV-2 Spikes and Virions by Single-Molecule Fluorescence

Authors:

Dasgupta, Souradip and Ray, Srijan and Srivatsav, Aswin T and DeVico, Anthony L and Ray, Krishanu

Journal:

ACS Omega (2025)

Genetically Programmable Self-Regenerating Bacterial Hydrogels

Authors:

Duraj-Thatte, Anna M and Courchesne, Noémie-Manuelle Dorval and Praveschotinunt, Pichet and Rutledge, Jarod and Lee, Yuhan and Karp, Jeffrey M and Joshi, Neel S

Journal:

Advanced Materials (2019): 1901826

References

View all 14 references: Citation Explorer

Purification of a Recombinant Polyhistidine-Tagged Glucosyltransferase Using Immobilized Metal-Affinity Chromatography (IMAC)

Authors:

de Costa F, Barber CJ, Pujara PT, Reed DW, Covello PS.

Journal:

Methods Mol Biol (2016): 91

New shuttle vector-based expression system to generate polyhistidine-tagged fusion proteins in Staphylococcus aureus and Escherichia coli

Authors:

Schwendener S, Perreten V.

Journal:

Appl Environ Microbiol (2015): 3243

Characterization of soluble RNA-dependent RNA polymerase from dengue virus serotype 2: The polyhistidine tag compromises the polymerase activity

Authors:

Kamkaew M, Chimnaronk S.

Journal:

Protein Expr Purif (2015): 43

Cellular uptake and in vivo distribution of polyhistidine peptides

Authors:

Iwasaki T, Tokuda Y, Kotake A, Okada H, Takeda S, Kawano T, Nakayama Y.

Journal:

J Control Release (2015): 115

Effects of the polyhistidine tag on kinetics and other properties of trehalose synthase from Deinococcus geothermalis

Authors:

Panek A, Pietrow O, Filipkowski P, Synowiecki J.

Journal:

Acta Biochim Pol (2013): 163

Physical properties

Molecular weight	1500.03
Solvent	Water

Spectral properties

Correction factor (260 nm)	0.02
Correction factor (280 nm)	0.03
Correction factor (482 nm)	0.009
Correction factor (565 nm)	0.09
Extinction coefficient (cm ^-1 M ^-1)	250000 ¹
Excitation (nm)	651
Emission (nm)	670
Quantum yield	0.27 ¹ , 0.4 ²

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
UNSPSC	12352200

Instrument settings

Gel Imager
Excitation	Red laser
Emission	700/50 nm

Telephone
Fax
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Technical Support	Contact us
Request quotation	Request
Purchase order	Send to sales@aatbio.com
Shipping	Standard overnight for United States, inquire for international

Page updated on October 12, 2025