FITC-Dextran (MW 10000)

• CAT Number: I018596
• CAS Number: 60842-46-8
• Molecular Weight: 10000.00(Average)
• Purity: ≥95%

FITC-Dextran (MW 10000) is a fluorescent probe for fluorescein isothiocyanate (FITC) dextran (Ex=495 nm; Em=525 nm). FITC-Dextran (MW 10000) can be used as a marker to reveal heat shock-induced cell damage and to study the early and late stages of apoptosis. FITC-Dextran (MW 10000) can also be used for cell permeability studies, such as blood-brain barrier permeability and determination of the extent of blood-brain barrier disruption[1][2][3].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Labeling of cells[1]:
For use with apoptotic HeLa cells and human peripheral blood mononuclear cells (PBMC) (viable HeLa and PBMC can not be stained by FITC-Dextran).
1. Incubate cells at 43.5°C for 1 hour and at 37°C for 8 hours to induce apoptosis.
2. Suspend the cells in 100 μL of medium, and mix in Q-prep tubes with 10 μL of propidium iodide (PI), 10 μL of FITC-Dextran (MW 10000) (the final concentration of PI and FITC-Dextran (MW 10000) is 7.5 μM and 1.13 μM, respectively).
3. Incubate cells for 25 min at room temperature in the dark.
4. Take the labeled cells with 3 mL of medium and centrifuge for 10 min at 500 g.
5. Take centrifuged cells with 1 mL of medium and use flow cytometry or fluorescence microscopy analyze (PI: Ex=500 nm, Em=600 nm; FITC-Dextran (MW 10000): Ex=495 nm, Em=525 nm).

Paracellular permeability measurement[4]
1. Add FITC-dextran (0.1 mg/mL) to the basal media in the transwell chamber.
2. Collect media from the transwell insert after 15 min.
3. Measure the fluorescence signal (Ex=485 nm, Em=538 nm).
4. Calculate FITC-dextran concentration based on fluorescence intensity.
5. Calculate permeability.
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
For intestinal barrier function assay[5]
1. Fast mice for 4 h.
2. Orally gavage mice with FITC-Dextran MW 10000 (0.6 mg/g).
3. Measure fluorescence intensity of plasma in 4 h (excitation nm/emission 520 nm).

Reference

[1]. Moumaris M, et al. Fluorescein isothiocyanate-dextran can track apoptosis and necrosis induced by heat shock of peripheral blood mononuclear cells and HeLa cells[J]. Open Biological Sciences Journal, 2015, 1(1).

[2]. Natarajan R, et al. Fluorescein Isothiocyanate (FITC)-Dextran Extravasation as a Measure of Blood-Brain Barrier Permeability. Curr Protoc Neurosci. 2017 Apr 10;79:9.58.1-9.58.15.
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[3]. Eriksson I, et al. Analysis of Lysosomal pH by Flow Cytometry Using FITC-Dextran Loaded Cells. Methods Mol Biol. 2017;1594:179-189.
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[4]. Okabayashi K, et al. Cdc42 activates paracellular transport in polarised submandibular gland cells. Arch Oral Biol. 2021 Dec;132:105276.
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[5]. Yu W, et al. ACE2 contributes to the maintenance of mouse epithelial barrier function. Biochem Biophys Res Commun. 2020 Dec 17;533(4):1276-1282.
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