El isotiocianato de fluoresceína ( FITC ) es un derivado de la fluoresceína que se utiliza en una amplia variedad de aplicaciones [1] [2], incluida la citometría de flujo . Descrito por primera vez en 1942, [3] FITC es la molécula de fluoresceína original funcionalizada con un grupo reactivo de isotiocianato (-N = C = S), reemplazando un átomo de hidrógeno en el anillo inferior de la estructura. Por lo general, está disponible como una mezcla de isómeros , 5-isotiocianato de fluoresceína (5-FITC) y 6-isotiocianato de fluoresceína (6-FITC). FITC es reactivo frente a nucleófilos, incluidos amina y sulfhidrilo. groups on proteins. It was synthesized by Robert Seiwald and Joseph Burckhalter in 1958. [4]
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Other names FITC 5-Isomer:
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3D model ( JSmol) | |||
ChEBI | |||
ChemSpider | |||
ECHA InfoCard | 100.043.810 | ||
MeSH | Fluorescein+isothiocyanate | ||
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CompTox Dashboard(EPA) | |||
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Properties | |||
C21H11NO5S | |||
Molar mass | 389.38 g·mol−1 | ||
Density | 1.542 g/mL | ||
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa). | |||
verify (what is ?) | |||
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A succinimidyl-ester functional group attached to the fluorescein core, creating "NHS-fluorescein", forms another common amine reactive derivative that has much greater specificity toward primary amines in the presence of other nucleophiles.
FITC has excitation and emission spectrum peak wavelengths of approximately 495 nm/519 nm,[5] giving it a green color. Like most fluorochromes, it is prone to photobleaching. Because of the problem with photobleaching, derivatives of fluorescein such as Alexa 488 and DyLight 488 have been tailored for various chemical and biological applications where greater photostability, higher fluorescence intensity, or different attachment groups are needed.
References
- ^ The TH; Feltkamp, T. E. (1970). "Conjugation of fluorescein isothiocyanate to antibodies: I. Experiments on the conditions of conjugation". Immunology. 18 (6): 865–873. PMC 1455721. PMID 5310665.
- ^ The TH; Feltkamp, T. E. (1970). "Conjugation of fluorescein isothiocyanate to antibodies: II. A reproducible method". Immunology. 18 (6): 875–881. PMC 1455722. PMID 4913804.
- ^ Coons AH, Creech HJ, Norman Jones R, Berliner E (1942). "The Demonstration of Pneumococcal Antigen in Tissues by the Use of Fluorescent Antibody". The Journal of Immunology. 45 (3): 159–170.
- ^ Riggs JL, Seiwald RJ, Burckhalter JH (1958). "Isothiocyanate Compounds as Fluorescent Labeling Agents for Immune Serum". The American Journal of Pathology. 34 (6): 1081–1097. PMC 1934794. PMID 13583098.
- ^ "FITC".