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腸道細(xì)菌及其毒素的移位可能是自身免疫性疾病發(fā)病的重要因素。這種猜測(cè)在一段時(shí)間內(nèi)被認(rèn)為是正確的。但是由于沒(méi)有合適的研究工具,因此沒(méi)有得到廣泛研究。事實(shí)上,研究表明壓力、手術(shù)和輕微胃腸道疾病等顯著增加了黏膜通透性。因此,這種情況下顯著增加了細(xì)菌從腸黏膜進(jìn)入人體的機(jī)率。
Chondrex公司提供FITC標(biāo)記的右旋糖苷及D-木糖檢測(cè)試劑盒用于評(píng)估體內(nèi)和體外大分子對(duì)半透膜的通透性。
通透性評(píng)估試劑盒
貨號(hào) | 中文名稱(chēng) | 英文名稱(chēng) | 規(guī)格 |
6601 | D-木糖檢測(cè)試劑盒 | D-Xylose Assay Kit | 1Kit |
通透性評(píng)估試劑-熒光標(biāo)記的右旋糖苷
貨號(hào) | 中文名稱(chēng) | 英文名稱(chēng) | 分子量 | 規(guī)格 |
4013 | FITC標(biāo)記的右旋糖苷 | FITC-Dextran | 4 kDa | 425 mg/ml x 5 ml |
4009 | FITC標(biāo)記的右旋糖苷 | FITC-Dextran | 40 kDa | 25 mg/ml x 5 ml |
4014 | FITC標(biāo)記的右旋糖苷 | FITC-Dextran | 70 kDa | 25 mg/ml x 5 ml |
一般來(lái)說(shuō),分子通過(guò)腸道進(jìn)入身體發(fā)生在細(xì)胞間或通過(guò)細(xì)胞進(jìn)入體內(nèi)。熒光標(biāo)記的右旋糖苷提供了簡(jiǎn)單,可靠的方法用于評(píng)估體外或體內(nèi)細(xì)胞旁通透性。另一種方法是,通過(guò)D-木糖測(cè)定跨細(xì)胞通透性,也就是說(shuō)通過(guò)D-木糖作為被動(dòng)載體介導(dǎo)小腸吸收。
FITC-標(biāo)記的右旋糖苷
一些研究表明壓力、手術(shù)和輕微胃腸道疾病等顯著增加了黏膜通透性。熒光標(biāo)記的右旋糖苷提供了簡(jiǎn)單,可靠的方法用于評(píng)估半透膜功能。關(guān)于FITC-標(biāo)記的右旋糖苷的分子量,( 40kDa),有研究表明在大腸桿菌中大多數(shù)蛋白分子量為25-120kDa,這表明具有生物學(xué)意義的蛋白質(zhì)在該范圍內(nèi)。因此Chondrex提供40kDa FITC-右旋糖苷作為相關(guān)大分子標(biāo)記物,用于體外或體內(nèi)半透膜通透性研究。
檢測(cè)流程:
1)飼喂前和實(shí)驗(yàn)過(guò)程中,禁食4小時(shí)。
2)口服40 kDa FITC-Dextran (10 mg/250 µl/mouse)。
3)3小時(shí)后,收集血漿。
4)1:2 (或更大)用PBS稀釋血漿
5)取50-100ul上去到96孔板。讀取熒光值:Ex: 490 nm Em: 530 nm
用PBS稀釋的FITC- 右旋糖苷繪制的標(biāo)準(zhǔn)曲線(xiàn)
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