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Biotin-4-fluorescein *CAS 1032732-74-3*

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Physical properties
Molecular weight644.69
SolventDMSO
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 yield0.79001, 0.952
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; Desiccated
UNSPSC12352200
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OverviewpdfSDSpdfProtocol


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

Excitation492 nm
Emission518 nm
Cutoff500 nm
Recommended plateSolid 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 mg0.5 mg1 mg5 mg10 mg
1 mM155.113 µL775.567 µL1.551 mL7.756 mL15.511 mL
5 mM31.023 µL155.113 µL310.227 µL1.551 mL3.102 mL
10 mM15.511 µL77.557 µL155.113 µL775.567 µL1.551 mL

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


Open in Advanced Spectrum Viewer
spectrum

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 yield0.79001, 0.952

Images


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

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
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
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
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
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
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
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
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
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
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