The Thermo Scientific FITC Antibody Labeling Kit is a multi-component kit containing a high-performance derivative of fluorescein dye, activated for easy and reliable labeling of antibodies, proteins and other molecules for use as fluorescent probes.
Features of the kit:
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Easy —convenient kit with FITC to label and purify antibody in about one hour
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Amine-specific labeling—the isothiocyanate variety of fluorescein efficiently labels antibodies and other purified proteins at primary amines (lysine side chains)
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Optimized kit procedure—following the standard protocol results in antibodies with excellent dye:protein ratios for optimum activity and fluorescence
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Single-use fluors—no need to weigh tiny amounts of powder; kits contain single-use vials of reagent
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Efficient purification—kits include purification resin and easy-to-use spin columns, ensuring rapid and efficient removal of non-reacted dye and excellent protein recovery
Properties of FITC:
• Alternative names: 5/6-FITC
• Chemical name: 5(6)-fluorescein isothiocyanate mixed isomer
• Molecular weight: 389.2
• Excitation source: 488 nm spectral line, argon-ion laser
• Excitation wavelength: 494 nm
• Emission wavelength: 518 nm
• Extinction coefficient: > 70,000 M
-1cm
-1• CAS #: 27072-45-3
• Purity: > 95% by HPLC
• Solubility: Soluble in aqueous buffers at pH > 6
• Reactive groups: Isothiocyanate, reacts with primary amines at pH 7.0 to 9.0
Applications:• Label antibodies for use as immunofluorescent probes
• Label oligonucleotides for hybridization probes
• Detect proteins in gels and on Western blots
Amine-reactive Fluorescein DyesFITC is the base fluorescein molecule functionalized with an
isothiocyanate reactive group (—N=C=S) at one of two hydrogen atoms on the bottom ring of the structure. This derivative is reactive towards primary amine groups on proteins, peptides and other biomolecules. NHS-fluorescein is activated with the
N-hydroxy-succinimidyl-ester (
NHS ester) functional group. Compared to FITC, the NHS-ester deriviative has greater specificity toward primary amines in the presence of other nucleophiles and results in a more stable linkage following labeling. Pierce Amine-reactive Fluorescein Dyes are mixtures of isomers with reactive groups attached at the 5- and 6-positions of the bottom ring. The properties of these isomers are indistinguishable in terms of excitation and emission spectra, and for protein applications there is no need to isolate a specific isomer.
Related ProductsNHS-Fluorescein (5/6-carboxyfluorescein succinimidyl ester), mixed isomerFITC (5/6-fluorescein isothiocyanate), mixed isomerPierce™ NHS-Fluorescein Antibody Labeling Kit