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- 保存条件:
4°C, protect from light
- 库存:
货期:1-2天
- 供应商:
MedChemExpress LLC
- CAS号:
60842-46-8
- 规格:
5 mg/10 mg/50 mg/100 mg
| 规格: | 5 mg | 产品价格: | ¥500.0 |
|---|---|---|---|
| 规格: | 10 mg | 产品价格: | ¥850.0 |
| 规格: | 50 mg | 产品价格: | ¥2500.0 |
| 规格: | 100 mg | 产品价格: | ¥4000.0 |
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FITC-Dextran (MW 10000)
CAS No. : 60842-46-8
MCE 国际站:FITC-Dextran (MW 10000)
产品活性:FITC-Dextran (MW 10000) 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 10000) 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 10000) 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。
研究领域:Others
作用靶点:Fluorescent Dye
In Vitro: Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Labeling of cells:
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
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.
In Vivo: 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
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).
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文献和实验天平准确称取所需的异硫氰酸荧光素粉末。③结合:边搅拌边将称取的荧光素渐渐加入抗体溶液中,避免将荧光素粘于三角烧瓶壁或搅拌玻棒上(大约在5~10min内加完),加完后,在4℃左右继续搅拌12~18h,通常将装置安放4℃冰箱或冰库中进行。④透析:结合完毕后,将标记的抗体溶液离心(2000r/min)10min,除去其中少量的沉淀物,装入透析袋中再置于烧杯中用0.01M pH7.2缓冲盐水透析(0~4℃)过夜。⑤过柱:取透析过夜的标记物,通过葡聚糖凝胶G-25或G-50柱,分离游离荧光素,收集标记的荧光抗体进行
(1)将标记完毕的荧光蛋白液装入一透析袋或玻璃纸袋内,液面稍留空隙,紧扎。 (2)浸入0.02mol/pH 7.1~7.4的PBS中(悬于大于标记物体积约50~100倍的PBS内),在4℃中透析,每日更换3~4次PBS,约5~7天,透析液中无荧即可(在荧光光源照射下)。 (三)葡聚糖凝胶G-50柱层析法 除游离荧光素可用2×46cm柱层析法,详细方法参阅第二章。加入荧光抗体15~18ml(按床体积的5%~10%加样),使其缓慢渗入柱内,待即将全部入柱时,加入PBS少许,关闭下口,停留30~
标记物体积约50~100倍的PBS内),在4℃中透析,每日更换3~4次PBS,约5~7天,透析液中无荧即可(在荧光光源照射下)。(三)葡聚糖凝胶G-50柱层析法除游离荧光素可用2×46cm柱层析法,详细方法参阅第二章。加入荧光抗体15~18ml(按床体积的5%~10%加样),使其缓慢渗入柱内,待即将全部入柱时,加入PBS少许,关闭下口,停留30~40min ,使游离荧光充分进入细筛孔中,然后再接通洗脱瓶开始滴入洗脱液。加入洗脱液一定量后,荧光抗体即向下移行,逐渐与存留于上端的游离荧光素之间拉开
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