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  • FITC-Dextran (MW 4000)
FITC-Dextran (MW 4000)的可视化放大

FITC-Dextran (MW 4000)

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)。

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  • 库存: 现货
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  • 货号: ajcf23898
  • CAS: 60842-46-8
  • 别名: 荧光素异硫氰酸酯-葡聚糖
  • 分子式:
  • 分子量: 4000.00(Average)
  • 纯度: >98%
  • 溶解度: Water: 16mg/mL
  • 储存: Store at 2-8°C, protect from light
  • 库存: 现货

Background

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]

Protocol

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。

  1. 五万个 bEnd.3 cells细胞接种于孔径为0.4 μm的transwell系统中,放置于24孔板中。
  2. 在transwell单层形成后,将transwell和下腔的培养基分别替换为条件培养基和对照培养基,孵育过夜。
  3. 上腔加入FITC-Dextran(0.1 mg/mL)。孵育20分钟后检测荧光穿透。
  4. 血管完整性评价为transwell和下腔之间的吸光度差异。

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]:

  1. 对肥胖和瘦弱小鼠肠道通透性的研究。小鼠按指定饮食喂养10-16周。

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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