FITC-Dextran (MW 4000) is a fluorescent probe for fluorescein isothiocyanate (FITC) dextran (Ex=495 nm; Em=525 nm). FITC-Dextran (MW 4000) can be used as a marker to reveal heat shock-induced cell damage and to study the early and late stages ofapoptosis. FITC-Dextran (MW 4000) 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 (HeLa cells for example)^[1]:
1. Suspend the cells in 100 μL of medium, and mix in Q-prep tubes with10 μLof propidium iodide (PI),10 μLof FITC-Dextran (MW 4000) (the final concentration of PI and FITC-Dextran (MW 4000) is7.5 μMand1.13 μM, respectively).
2. Incubate cells for25 minat room temperature in the dark.
3. Take the labeled cells with3 mLof medium and centrifuge for10 minat 500g.
4. Take centrifuged cells with1 mLof medium and use flow cytometry or fluorescence microscopy analyze (PI: Ex=500 nm, Em=600 nm; FITC-Dextran (MW 4000): Ex=495 nm, Em=525 nm).

4000.00(Average)

Solid

60842-46-8

荧光素异硫氰酸酯-葡聚糖(MW 4000)

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

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

DMSO : 16.67 mg/mL(Need ultrasonic)

H₂O : 16.67 mg/mL(Need ultrasonic)

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照In Vitro方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： PBS

  Solubility: 100 mg/mL (Infinity mM); Clear solution; Need ultrasonic
• 2.

  请依序添加每种溶剂： 10% DMSO    40%PEG300   5%Tween-80   45% saline

  Solubility: 1.67 mg/mL (Infinity mM); Clear solution; Need ultrasonic

  此方案可获得 1.67 mg/mL (Infinity mM) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 16.7 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH2O 中，得到澄清透明的生理盐水溶液
• 3.

  请依序添加每种溶剂： 10% DMSO    90% (20%SBE-β-CDin saline)

  Solubility: 1.67 mg/mL (Infinity mM); Suspended solution; Need ultrasonic

  此方案可获得 1.67 mg/mL (Infinity mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  以 1 mL 工作液为例，取 100 μL 16.7 mg/mL 的澄清 DMSO 储备液加到 900 μL20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 4.

  请依序添加每种溶剂： 10% DMSO    90%corn oil

  Solubility: 1.67 mg/mL (Infinity mM); Clear solution; Need ultrasonic

  此方案可获得 1.67 mg/mL (Infinity mM) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 16.7 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• 
  Description: FITC-Dextran MW 4000 can be used to explore the cellular uptake with green fluorescence.
  Method: For cellular uptake monitoring.
  1. Incubate cells in glass dishes (37℃; overnight).
  2. Incubate cells with FITC-Dextran MW 4000 (5 μg/mL; 4 h).
  3. Wash cells for twice with PBS and finally fix cells with 4% PFA (30 min; dark).
  4. Use a confocal laser scanning microscopy (Zeiss 880) for image.
• 
  Description: FITC-Dextran MW 4000 can be used to detect intestinal barrier function in vivo.
  Method: For assess intestinal barrier function.
  1. Fast mice for 4 hours.
  2. Orally gavage mice with FITC-Dextran MW 4000 (0.6 mg/g).
  3. Measure fluorescence intensity of plasma in 4 hours.
• 
  Description: FITC-Dextran MW 4000 can be used for labeling the microvessels in the lesion area of rat brain to measure microvessel density.
  Method: For vivo labeling.
  1. Anesthetize rats first.
  2. Inject FITC-dextran FITC-Dextran MW 4000 (50 mg/kg) in tail vein of rats.
  3. Remove brain tissues 1 min after injection, use 4% paraformaldehyde (4℃; 24 h) to fix tissues and cut tissues into 150 μm-thick sections.
  4. Use a laser scanning confocal microscope (Leica, Solms, Germany) for image.