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Cy5 biotin conjugate

This bifunctional Cy5 biotin conjugate demonstrates excellent avidin binding and strong fluorescence. It has avidin-binding properties similar to native biotin in terms of high affinity, fast association, and non-cooperative binding to avidin and streptavidin tetramers. Cy5 Biotin can be used for detecting and quantifying biotin binding sites of avidin, streptavidin or neutravidin. This reagent overcomes major shortcomings of commonly used Biotin-4-fluorescein, e.g., poor solubility in aqueous media and pH dependent absorbance and emission. Cy5 Biotin is a water-soluble reagent and its red fluorescence is pH independent from pH 4 to pH 10. A flexible PEG spacer between biotin moiety and fluorescent tag minimize steric hindrance involved in binding to avidin, streptavidin or neutravidin.

Example protocol

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of Cy5 biotin conjugate 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 mM73.865 µL369.325 µL738.651 µL3.693 mL7.387 mL
5 mM14.773 µL73.865 µL147.73 µL738.651 µL1.477 mL
10 mM7.387 µL36.933 µL73.865 µL369.325 µL738.651 µL

Molarity calculator

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

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum

Citations

View all 27 citations: Citation Explorer
Streptavidin cooperative allosterism upon binding biotin observed by differential changes in intrinsic fluorescence
Authors: Waner, M. J., Hiznay, J. M., Mustovich, A. T., Patton, W., Ponyik, C., Mascotti, D. P.
Journal: Biochem Biophys Rep (2019): 127-131
Biotin-tagged fluorescent sensor to visualize 'mobile' Zn(2+) in cancer cells
Authors: Fang, L., Trigiante, G., Kousseff, C. J., Crespo-Otero, R., Philpott, M. P., Watkinson, M.
Journal: Chem Commun (Camb) (2018): 9619-9622
Synthesis of Two-Photon Active Tricomponent Fluorescent Probe for Distinguishment of Biotin Receptor Positive and Negative Cells and Imaging 3D-Spheroid
Authors: Pal, K., Sharma, A., Koner, A. L.
Journal: Org Lett (2018): 6425-6429
A green fluorescent protein-based assay for high-throughput ligand-binding studies of a mycobacterial biotin protein ligase
Authors: Bond, T. E. H., Sorenson, A. E., Schaeffer, P. M.
Journal: Microbiol Res (2017): 35-39
Labeling of Cytoskeletal Proteins in Living Cells Using Biotin Ligase Carrying a Fluorescent Protein
Authors: Nishi, S., Yamamoto, C., Yoneda, S., Sueda, S.
Journal: Anal Sci (2017): 897-902

References

View all 81 references: Citation Explorer
A low-cost, high-performance system for fluorescence lateral flow assays
Authors: Lee, Linda G and Nordman, Eric S and Johnson, Martin D and Oldham, Mark F
Journal: Biosensors (2013): 360--373
Implementation of P22 viral capsids as nanoplatforms
Authors: Kang, Sebyung and Uchida, Masaki and O'Neil, Alison and Li, Rui and Prevelige, Peter E and Douglas, Trevor
Journal: Biomacromolecules (2010): 2804--2809
Evaluation of a new biotin-DOTA conjugate for pretargeted antibody-guided radioimmunotherapy (PAGRIT((R)))
Authors: Urbano N, Papi S, Ginanneschi M, De Santis R, Pace S, Lindstedt R, Ferrari L, Choi S, Paganelli G, Chinol M.
Journal: Eur J Nucl Med Mol Imaging. (2006)
A biotin-protein bond with stability in plasma
Authors: Bogusiewicz A, Mock NI, Mock DM.
Journal: Anal Biochem (2005): 98
Instability of the biotin-protein bond in human plasma
Authors: Bogusiewicz A, Mock NI, Mock DM.
Journal: Anal Biochem (2004): 156
Page updated on December 17, 2024

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Catalog Number3100
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Physical properties

Molecular weight

1353.82

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)

2500001

Excitation (nm)

651

Emission (nm)

670

Quantum yield

0.271, 0.42

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200

Platform

Other instruments

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