Biotin-4-fluorescein *CAS 1032732-74-3*
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Physical properties
Molecular weight | 644.69 |
Solvent | DMSO |
Spectral properties
Absorbance (nm) | 487 |
Correction Factor (260 nm) | 0.32 |
Correction Factor (280 nm) | 0.35 |
Extinction coefficient (cm -1 M -1) | 800001 |
Excitation (nm) | 498 |
Emission (nm) | 517 |
Quantum yield | 0.79001, 0.952 |
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; Desiccated |
UNSPSC | 12352200 |
Related products
Overview | SDSProtocol |
See also: Biotin and Streptavidin, Fluoresceins
CAS 1032732-74-3 | Molecular weight 644.69 | Absorbance (nm) 487 | Correction Factor (260 nm) 0.32 | Correction Factor (280 nm) 0.35 | Extinction coefficient (cm -1 M -1) 800001 | Excitation (nm) 498 | Emission (nm) 517 | Quantum yield 0.79001, 0.952 |
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.
Platform
Fluorescence microplate reader
Excitation | 492 nm |
Emission | 518 nm |
Cutoff | 500 nm |
Recommended plate | Solid black |
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
Open in Advanced Spectrum Viewer
Spectral properties
Absorbance (nm) | 487 |
Correction Factor (260 nm) | 0.32 |
Correction Factor (280 nm) | 0.35 |
Extinction coefficient (cm -1 M -1) | 800001 |
Excitation (nm) | 498 |
Emission (nm) | 517 |
Quantum yield | 0.79001, 0.952 |
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
Synthesis of adenosine derivatives as transcription initiators and preparation of 5' fluorescein- and biotin-labeled RNA through one-step in vitro transcription
Authors: Huang F, Wang G, Coleman T, Li N.
Journal: Rna (2003): 1562
Authors: Huang F, Wang G, Coleman T, Li N.
Journal: Rna (2003): 1562
A novel intravascular drug delivery method using endothelial biotinylation and avidin-biotin binding
Authors: Hoya K, Guterman LR, Miskolczi L, Hopkins LN.
Journal: Drug Deliv (2001): 215
Authors: Hoya K, Guterman LR, Miskolczi L, Hopkins LN.
Journal: Drug Deliv (2001): 215
Organelle pH studies using targeted avidin and fluorescein-biotin
Authors: Wu MM, Llopis J, Adams S, McCaffery JM, Kulomaa MS, Machen TE, Moore HP, Tsien RY.
Journal: Chem Biol (2000): 197
Authors: Wu MM, Llopis J, Adams S, McCaffery JM, Kulomaa MS, Machen TE, Moore HP, Tsien RY.
Journal: Chem Biol (2000): 197
Accurate measurement of avidin and streptavidin in crude biofluids with a new, optimized biotin-fluorescein conjugate
Authors: Kada G, Falk H, Gruber HJ.
Journal: Biochim Biophys Acta (1999): 33
Authors: Kada G, Falk H, Gruber HJ.
Journal: Biochim Biophys Acta (1999): 33
Rapid estimation of avidin and streptavidin by fluorescence quenching or fluorescence polarization
Authors: Kada G, Kaiser K, Falk H, Gruber HJ.
Journal: Biochim Biophys Acta (1999): 44
Authors: Kada G, Kaiser K, Falk H, Gruber HJ.
Journal: Biochim Biophys Acta (1999): 44