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FITC-Dextran (MW 4000) 是一种异硫氰酸荧光素 (FITC) 葡聚糖荧光探针 (Ex=495 nm; Em=525 nm)。
FITC-Dextran (MW 4000) (FD4) is a large molecule of dextran (MW ~4000) attached to a fluorescein isothiocyanate tag, which makes it fluorescent (Ex=495 nm; Em=525 nm) [1-2].
FITC-Dextran (MW 4000) can be used for the study of cell permeability in bEnd.3 cells in vitro(0.1 mg/mL) and blood-brain barrier (BBB) permeability [3-4]. FITC-Dextran (MW 4000) can also be used as a marker to reveal cell damage caused by heat shock and to study the early and late stages of apoptosis[5].
FITC-Dextran (MW 4000) (0.6mg/g; i.g) can also be used to study intestinal barrier function [6].FITC-Dextran (MW 4000) (0.4mg/kg; i.p; 3days) can also be used to study blood-brain barrier permeability in male sprague dawley rats[7].
参考文献:
[1]. Eriksson I, ?llinger K, et,al. Analysis of Lysosomal pH by Flow Cytometry Using FITC-Dextran Loaded Cells. Methods Mol Biol. 2017;1594:179-189. doi: 10.1007/978-1-4939-6934-0_11. PMID: 28456983.
[2]. Tsuneoka Y, Nishimura T, et,al. Fluorescein Permeability of the Blood-Brain Barrier Is Enhanced in Juvenile- but Not Young Adult-Onset Type 1 Diabetes in Rats. Biol Pharm Bull. 2021;44(8):1088-1092. doi: 10.1248/bpb.b21-00248. PMID: 34334494.
[3]. Wu S, Wang J et,al. Programmed cell death 10 increased blood-brain barrier permeability through HMGB1/TLR4 mediated downregulation of endothelial ZO-1 in glioblastoma. Cell Signal. 2023 Jul;107:110683. doi: 10.1016/j.cellsig.2023.110683. Epub 2023 Apr 17. PMID: 37075875.
[4]. Natarajan R, Northrop N 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. doi: 10.1002/cpns.25. PMID: 28398646; PMCID: PMC5470084.
[5]. 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).
[6]. Yu W, Ou X, et,al. ACE2 contributes to the maintenance of mouse epithelial barrier function. Biochem Biophys Res Commun. 2020 Dec 17;533(4):1276-1282. doi: 10.1016/j.bbrc.2020.10.002. Epub 2020 Oct 21. PMID: 33097186; PMCID: PMC7576438.
[7]. Bommana MM, Kirthivasan B, et,al. In vivo brain microdialysis to evaluate FITC-dextran encapsulated immunopegylated nanoparticles. Drug Deliv. 2012 Aug;19(6):298-306. doi: 10.3109/10717544.2012.714812. Epub 2012 Aug 28. PMID: 22928708.
FITC-Dextran (MW 4000) (FD4)是一种大分子葡聚糖(MW ~4000)附着在异硫氰酸荧光素标签上,具有荧光性(Ex=495 nm; Em=525 nm) [1-2]。
FITC-Dextran (MW 4000) (0.1 mg/mL)可用于bEnd.3细胞通透性以及血脑屏障通透性研究[3-4]。FITC-Dextran (MW 4000)也可作为热休克细胞损伤的标志物,研究细胞凋亡的早期和晚期[5]。
FITC-Dextran (MW 4000) (0.6mg/g;i.g)也可用于研究肠屏障功能[6]。FITC-Dextran (MW 4000) (0.4mg/kg;i.p;3days)也可用于雄性大鼠血脑屏障通透性的研究[7]。
FITC-Dextra体外血脑屏障通透性测定[1]: |
体外血脑屏障通透性研究采用transwell法。
|
细胞标记实验[2]: |
适用于正在发生凋亡的HeLa细胞和人外周血单个核细胞(PBMC)。 1. 43.5℃孵育1h,37℃孵育8h,诱导细胞凋亡。 2. 将细胞悬浮在100 μL的培养基中,在Q-prep管与10 μL的propidium iodide (PI),10 μL的FITC-Dextran (MW 4000)混合,PI和FITC-Dextran (MW 4000)的最终浓度分别为7.5 μM和1.13 μM。 3. 将细胞在室温黑暗条件下培养25分钟。 4. 取3 mL培养基的标记细胞,500 g离心10分钟。 5. 取离心细胞,加入1 mL培养基,用流式细胞术或荧光显微镜分析。 |
体内肠通透性试验方案[3]: |
将小鼠体重分为两组,LBM剂量组和BW剂量组(瘦体重组和肥胖体重组)。 在研究结束时,小鼠为18-24周大。 2.??? 在研究当天,小鼠禁食4小时,然后按随机顺序口服FITC-Dextran (MW 4000) (溶解在浓度为125 mg/ml的PBS中)(600 mg/kg)。 3.??? 给药2小时后,用异氟烷麻醉小鼠(诱导5%,维持2%异氟烷,0.7 L/min N2O, 0.3 L/min O2),在K3-EDTA-coated管中取眶后血,然后颈椎脱位。 4.??? 样品离心(4℃,7 min, 8000 g),血浆收集于透明的Eppendorf管中。 5.?? PBS给药小鼠的血浆用于标准曲线。用分光光度计分析血浆中FITC-Dextran浓度。 This protocol only provides a guideline, and should be modified according to your specific needs. |
参考文献: [1]. Wu S, Wang J, et,al. Programmed cell death 10 increased blood-brain barrier permeability through HMGB1/TLR4 mediated downregulation of endothelial ZO-1 in glioblastoma. Cell Signal. 2023 Jul;107:110683. doi: 10.1016/j.cellsig.2023.110683. Epub 2023 Apr 17. PMID: 37075875. [2]. 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). [3]. Voetmann LM, Rolin B, et al. The intestinal permeability marker FITC-dextran 4kDa should be dosed according to lean body mass in obese mice. Nutr Diabetes. 2023 Jan 5;13(1):1. doi: 10.1038/s41387-022-00230-2. PMID: 36604407; PMCID: PMC9816099. |
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