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AAT Bioquest

HIS Lite™ Cy5 Bis NTA Chelator

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Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
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Physical properties
Molecular weight1203.34
SolventDMSO
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)2500001
Excitation (nm)651
Emission (nm)670
Quantum yield0.271, 0.42
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
1203.34
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)
2500001
Excitation (nm)
651
Emission (nm)
670
Quantum yield
0.271, 0.42
Cy5 Bis NTA-Ni Chelator can be applied either to an SDS-PAGE gel or Western blot membrane for fluorescence imaging or used in living cells in combination with the addition of a heavy metal ion such as Ni2+. Detection with Cy5 Bis NTA-Ni complex requires less incubation time than for protein-antibody binding. No secondary reaction is required since Ni-NTA complex is directly conjugated to the Cy5 fluorophores. Cy5 demonstrates strong fluorescent signals at ~670 nm with a common red laser excitation (e.g., He-Ne laser or 647 nm red laser). 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. AAT Bioquest also offers HIS Lite™ Cy5 Bis 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.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of HIS Lite™ Cy5 Bis 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 mM83.102 µL415.51 µL831.02 µL4.155 mL8.31 mL
5 mM16.62 µL83.102 µL166.204 µL831.02 µL1.662 mL
10 mM8.31 µL41.551 µL83.102 µL415.51 µL831.02 µL

Molarity calculator

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Spectrum


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spectrum

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)2500001
Excitation (nm)651
Emission (nm)670
Quantum yield0.271, 0.42

Product Family


NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yieldCorrection Factor (260 nm)Correction Factor (280 nm)Correction Factor (482 nm)Correction Factor (565 nm)
HIS Lite™ Cy5 Tris NTA Chelator65167025000010.271, 0.420.020.030.0090.09
HIS Lite™ Cy3 Bis NTA Chelator55556915000010.1510.070.073--

Images


References


View all 9 references: Citation Explorer
Expression and purification of recombinant arginine decarboxylase (speA) from Escherichia coli.
Authors: Song, Jiaping and Zhou, Chuanwen and Liu, Rui and Wu, Xudong and Wu, Di and Hu, Xiaojian and Ding, Yu
Journal: Molecular biology reports (2010): 1823-9
[Isolation, prokaryotic expression and activity analysis of thymidylate kinase (tmk) gene from Phytoplasma of wheat blue dwarf].
Authors: Li, Bei and Ji, Lingling and Wu, Yunfeng and Hao, Xing'an
Journal: Wei sheng wu xue bao = Acta microbiologica Sinica (2008): 739-44
Bacterial elongation factors EF-Tu, their mutants, chimeric forms, and domains: isolation and purification.
Authors: Jonák, J
Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (2007): 141-53
[Displaying polyhistidine peptide on the cell surface of Bacillus thuringiensis by S-layer protein].
Authors: Wang, Li and Sun, Ming and Yu, Zi Niu
Journal: Shi yan sheng wu xue bao (2003): 476-81
Lack of antitumor activity of recombinant endostatin in a human neuroblastoma xenograft model.
Authors: Jouanneau, E and Alberti, L and Nejjari, M and Treilleux, I and Vilgrain, I and Duc, A and Combaret, V and Favrot, M and Leboulch, P and Bachelot, T
Journal: Journal of neuro-oncology (2001): 11-8
Expression and functional characterization of a pHis-tagged human bradykinin B2 receptor in COS-7 cells.
Authors: Müller, S and Adomeit, A and Kaufmann, R and Appelhans, H and Passow, H and Reissmann, S and Liebmann, C
Journal: Biological chemistry (2000): 343-7
The cloning, expression and purification of cervine interleukin 2.
Authors: Lockhart, E and Griffin, J F and Chinn, N and Lavallie, E and Buchan, G
Journal: Cytokine (1996): 603-12
Expression of biologically active C3a as fusion proteins.
Authors: Fukuoka, Y and Tachibana, T and Yasui, A
Journal: Immunology letters (1993): 153-8
A metal-chelating piezoelectric sensor chip for direct detection and oriented immobilization of polyHis-tagged proteins.
Authors: Chen, Hsiu-Mei and Wang, Wei-Cheng and Chen, Sheng-Horng
Journal: Biotechnology progress : 1237-44