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Biotin-4-fluorescein *CAS 1032732-74-3*
This bifunctional biotin-fluorescein conjugate demonstrates better binding and stronger fluorescence than biotin fluorescein. It has similar avidin-binding properties in terms of high affinity, fast association, and non-cooperative binding to avidin and streptavidin tetramers. These exceptional properties are attributed to the small size/length of the new ligand since all larger/longer biotin derivatives are known for their mutual steric hindrance and anti-cooperative binding in 4:1 complexes with avidin and streptavidin tetramers. Specific binding of this biotin-fluorescein conjugate towards avidin and streptavidin is accompanied by 84-88% quenching of ligand fluorescence. It is used for the quantitation of biotin-binding sites. Both the fluorescence and absorbance of biotin-4-fluorescein are quenched upon binding to one of the four biotin-binding sites of streptavidin, avidin, or the streptavidin conjugates of fluorescent dyes or enzymes. As a result, when a known concentration of biotin-4-fluorescein is added to a known amount of streptavidin, one can estimate the number of biotin-binding sites.
Example protocol
AT A GLANCE
Important notes
It is important to store at <-15 °C and should be stored in cool, dark place.
It can be used within 12 months from the date of receipt.
PREPARATION OF WORKING SOLUTION
Biotin-4-fluorescein working solution:
Add appropriate amount of DMSO to make Biotin-4-fluorescein working solution.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of Biotin-4-fluorescein *CAS 1032732-74-3* 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 | 155.113 µL | 775.567 µL | 1.551 mL | 7.756 mL | 15.511 mL |
| 5 mM | 31.023 µL | 155.113 µL | 310.227 µL | 1.551 mL | 3.102 mL |
| 10 mM | 15.511 µL | 77.557 µL | 155.113 µL | 775.567 µL | 1.551 mL |
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 1 citations: Citation Explorer
Nanobody production can be simplified by direct secretion from Escherichia coli
Authors: Iwaki, Takayuki and Hara, Kimiko and Umemura, Kazuo
Journal: Protein Expression and Purification (2020): 105607
Authors: Iwaki, Takayuki and Hara, Kimiko and Umemura, Kazuo
Journal: Protein Expression and Purification (2020): 105607
References
View all 17 references: Citation Explorer
Engineered single-chain dimeric streptavidins with an unexpected strong preference for biotin-4-fluorescein
Authors: Aslan FM, Yu Y, Mohr SC, Cantor CR.
Journal: Proc Natl Acad Sci U S A (2005): 8507
Authors: Aslan FM, Yu Y, Mohr SC, Cantor CR.
Journal: Proc Natl Acad Sci U S A (2005): 8507
Dynamics of fluorescence dequenching of ostrich-quenched fluorescein biotin: a multifunctional quantitative assay for biotin
Authors: Wu Y, Simons PC, Lopez GP, Sklar LA, Bur and a T., undefined
Journal: Anal Biochem (2005): 221
Authors: Wu Y, Simons PC, Lopez GP, Sklar LA, Bur and a T., undefined
Journal: Anal Biochem (2005): 221
Preparation and characterisation of antibody modified gelatin nanoparticles as drug carrier system for uptake in lymphocytes
Authors: Balthasar S, Michaelis K, Dinauer N, von Briesen H, Kreuter J, Langer K.
Journal: Biomaterials (2005): 2723
Authors: Balthasar S, Michaelis K, Dinauer N, von Briesen H, Kreuter J, Langer K.
Journal: Biomaterials (2005): 2723
Electrophoretic behavior of streptavidin complexed to a biotinylated probe: a functional screening assay for biotin-binding proteins
Authors: Humbert N, Zocchi A, Ward TR.
Journal: Electrophoresis (2005): 47
Authors: Humbert N, Zocchi A, Ward TR.
Journal: Electrophoresis (2005): 47
Controlled surface functionalization of silica nanospheres by covalent conjugation reactions and preparation of high density streptavidin nanoparticles
Authors: Schiestel T, Brunner H, Tovar GE.
Journal: J Nanosci Nanotechnol (2004): 504
Authors: Schiestel T, Brunner H, Tovar GE.
Journal: J Nanosci Nanotechnol (2004): 504
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