FITC-Dextran (MW 40000)

国际站：FITC-Dextran (MW 40000)

CAS：60842-46-8

品牌：MedChemExpress (MCE)

存储条件：4°C, protect from light *In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

生物活性：FITC-Dextran (MW 40000) 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 400000) 可作为一种标记物来揭示热休克引起的细胞损伤，并研究细胞凋亡的早期和晚期阶段。FITC-Dextran (MW 400000) 还可用于细胞渗透性的研究，如血脑屏障通透性以及血脑屏障破坏程度的测定。

体外：Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).细胞标记[1]:适用于正在发生凋亡的 HeLa 细胞和人外周血单个核细胞 (PBMC) (活的 HeLa 和 PBMC 不被 FITC-Dextran 染色)。1. 43.5°C 孵育 1 h，37°C 孵育 8 h，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 40000) 混合，PI 和 FITC-Dextran (MW 40000) 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 (PI: Ex=500 nm, Em=600 nm; FITC-Dextran (MW 40000): Ex=495 nm, Em=525 nm)。 细胞旁渗透性检测[4]1. 在 transwell 室的基础培养基中加入 FITC-Dextran (0.1 mg/mL)。2. 在 15 分钟后从 transwell 插入器中收集培养基。3. 测量荧光信号 (Ex=485 nm, Em=538 nm)。4. 根据荧光强度计算 FITC-Dextran 浓度。5. 计算渗透率。 MCE has not independently confirmed the accuracy of these methods. They are for reference only.

体内：Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).肠屏障功能测定[5]1. 小鼠饥饿处理 4 小时。2. FITC-Dextran MW 40000 灌胃小鼠 (0.6 mg/g)。3. 在 4 h 内测量荧光强度 (Ex nm/Em 520 nm)。 MCE has not independently confirmed the accuracy of these methods. They are for reference only.

参考文献：[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.[3]. Eriksson I, et al. Analysis of Lysosomal pH by Flow Cytometry Using FITC-Dextran Loaded Cells. Methods Mol Biol. 2017;1594:179-189.[4]. Okabayashi K, et al. Cdc42 activates paracellular transport in polarised submandibular gland cells. Arch Oral Biol. 2021 Dec;132:105276.[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.