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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 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
| 1 mM | 73.865 µL | 369.325 µL | 738.651 µL | 3.693 mL | 7.387 mL |
| 5 mM | 14.773 µL | 73.865 µL | 147.73 µL | 738.651 µL | 1.477 mL |
| 10 mM | 7.387 µL | 36.933 µL | 73.865 µL | 369.325 µL | 738.651 µL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
| Mass (Calculate) | Molecular weight | Volume (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
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
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
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
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
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
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
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)
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
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
Authors: Bogusiewicz A, Mock NI, Mock DM.
Journal: Anal Biochem (2004): 156
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