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Dextran FITC, dextran fluorescein, (dextran FITC)
CAS NO: 60842-46-8
English Name:dextran (3,6'dihydroxy-3-oxospiro (isobenzofuran-1 (3H), 9 - [9h] xanthen] - 5 (or 6] - YL] carbamothioate, average MW of approximation
Molecular weight: 4000, 5000, 6000, 7000, 9000, 10000, 20000, 40000, 70000, etc., up to 2000K.
Solubility: soluble in water or salt solution, also soluble in DMSO, formamide and some other polar organic solvents, insoluble in lower fatty alcohols, acetone, chloroform and dimethylformamide (DMF)
Properties: White / off white solid powder or viscous liquid based on different molecular weights.
Stability: keep sealed and dry and refrigerated to avoid repeated freezing and thawing.
FITC dextran (FITC dextran) is a synthetic probe labeled with fluorescein, using dextran of different molecular weight prepared by restriction hydrolysis and fractionation purification of natural dextran b512f as substrate. The fluorescein group is attached to dextran by a stable thiocarbamoyl bond, and this labeling step does not cause any degradation of dextran. Each glucose unit in FITC dextran contains 0.002 ~ 0.008 mol FITC. This low level of substitution ensures the minimum structural change of dextran, which is the most basic requirement for permeability research.
FITC dextran (FITC dextran) is widely used in microcirculation and cell fitness studies, and is detected by fluorescence microscopy. It can also be used to study the porosity of plant cell wall and capillary permeability. Plasma proteins do not bind FITC dextran.
FITC dextran can also be used for cardiovascular, microcirculation, perfusion, cell monolayer and cell membrane permeability studies. As a fluorescent flow tracer compound, it can support the determination of blood flow, membrane damage, vascular drainage and renal clearance. Fitc-dextran40 may be used to study endocytosis. It may also be used to study the porosity of endothelial single cell layer and the movement from the top to the basement membrane within the cell layer.
This material is a fluorescein derivative labeled with dextran with an average molecular weight of about 4kDa, i.e. fluorescein isothiocyanate dextran 4 (FITC dextran, FD4). It is provided in the form of lyophilized powder and is easily soluble in water. The maximum excitation and emission wavelengths are 493 and 520nm respectively.
In general, the stability of FITC dextrans in vitro and in vivo is excellent according to the stability determination of FITC dextrans under various solution systems and temperatures. There may be a risk of hydrolysis of fluorescein only when pH > 9 and rising temperature. FITC dextrans in rabbit plasma, muscle homogenate, liver homogenate and urine remained stable at 37 ° C for at least 3 days, with no change in molecular weight and no release of fluorescein groups. 6% trichloroacetic acid in FITC dextrans at room temperature for 3 days. [literature source: P. kurtzhals, C. Larsen and M. Johansen, high performance size exclusion chromatographic procedure for the determination of fluorescein YL isothiocyanate ranges of variable molecular masses in biological media. J Chromatogr, 491 (1989), 117-127.]
Hydrolysis of thiocarbamoyl bond produces 4 - or 5-aminofluorescein, which can be detected quantitatively by HPLC immediately. Unpublished research data showed that when the high-pressure treated FITC dextran 70 solution was placed at 8-50 ° C for 5 months, it was found that only at 50 ° C, a slight (1%) increase in the content of free amino fluorescein could be observed. High pressure treatment alone caused 2.7% free amino fluorescein production.
Other unpublished studies show that FITC dextran can be stably stored for up to 1 year in pH4 solution at 35 ℃. However, the degradation of dextran may be observed when the thiocarbamoyl bond is stable for 30min at pH4 and 80 ℃. At pH 9, a certain amount (24%) of fluorescence attenuation will occur when stored at 35 ℃ for more than one month.
Some studies have demonstrated the in vivo stability of FITC dextrans in continuous experiments (1-6 days). [source: n. thorball, FITC dextran traces in microcirculatory and capability studies using combined fluorescence stereo microscopy, fluorescence stereo microscopy and electron microscopy, histochemistry, 71 (1981), 209-233.]
matters needing attention
FITC dextrans is easily soluble in water or salt solution, and the mother liquor of appropriate concentration can be prepared according to the actual concentration.
High pressure sterilization of FITC dextrans solution may lead to the release of a small amount of Aminofluorescein. If sterility is required, it is recommended to use filtration to remove bacteria.
The dextran of Yuyu pharmaceutical can connect different activating groups such as NH2 / mal / COOH, and can also provide some fluorescently labeled dextran such as FITC / rhodamine.
In order to ensure the stability of the product, please note that the package can only be opened for use after constant temperature drying!
Note: products with different group substitution degree can be customized and synthesized according to customer needs!
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