enzo life sciences公司的efluxx-id®多藥耐藥性分析試劑盒可實(shí)現(xiàn)對三個臨床相關(guān)的abc轉(zhuǎn)運(yùn)蛋白mdr1 (p-glycoprotein), mrp1/2和bcrp的功能檢測。試劑盒采用疏水性的非熒光化合物，很容易穿透細(xì)胞膜，細(xì)胞內(nèi)脂酶將其水解為親水性的熒光染料。除非abc轉(zhuǎn)運(yùn)蛋白通過主動運(yùn)輸將其泵出胞外，否則產(chǎn)生的熒光染料將包裹在細(xì)胞內(nèi)。因此如果細(xì)胞對藥物有抗性作用，熒光強(qiáng)度就會減弱。efluxx-id®多藥耐藥性分析試劑盒是目前少有可同時監(jiān)測三種主要的abc轉(zhuǎn)運(yùn)蛋白并能夠分析單泵活性的試劑盒。
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efluxx-id®多藥耐藥性染料與其他熒光染料的兼容性
產(chǎn)品信息
貨號
產(chǎn)品名稱
規(guī)格
enz-51029-k100
efluxx-id®green multidrug resistance assay kit
1*1kit(100 assays for flow cytometry)
enz-51030-k100
efluxx-id®gold multidrug resistance assay kit
1*1kit(100 assays for flow cytometry)
產(chǎn)品特點(diǎn)
●可檢測與3種abc轉(zhuǎn)運(yùn)蛋白活性相關(guān)的多藥耐藥性
●通過單一專有染料定量檢測活細(xì)胞中mdr活性，并以mdr活性因子（maf）表征
●試劑盒內(nèi)含有已知的mdr1、mrp1/2和bcrp特異性抑制劑
●操作簡單，無需洗滌，1h內(nèi)可獲得結(jié)果
●可檢測calcein am無法檢測到的bcrp蛋白活性
●有綠色和金色兩種熒光染料可供選擇
●兩種染料均可與表達(dá)gfp的細(xì)胞系或其他cellestial®染料同時使用
實(shí)例分析
●efluxx-id®green 490/514nm ex/em和efluxx-id®gold 530/570nm ex/em試劑的光譜特性可與其他常見熒光染料進(jìn)行多重檢測。
●使用efluxx-id®green和efluxx-id®gold染料在cho k1細(xì)胞中評估已知抑制劑對abc轉(zhuǎn)運(yùn)蛋白活性的分析
將細(xì)胞與試劑盒中含有的mdr通用抑制劑（最左列）或轉(zhuǎn)運(yùn)蛋白特異性抑制劑在37°c下孵育5 min。隨后在37°c下用指-定的染料染色細(xì)胞30 min，并立即通過流式細(xì)胞術(shù)進(jìn)行分析。所用抑制劑：5 µm cyclosporin a（mdr通用抑制劑）、20 µm verapamil（p-gp特異性抑制劑）、0.05 mm mk-571（mrp特異性抑制劑）、0.05 mm novobiocin（bcrp特異性抑制劑）。
●三種主要abc轉(zhuǎn)運(yùn)蛋白的活性分析
使用efluxx-id®green（上）、gold（中）或calcein am（下）染料，通過流式細(xì)胞術(shù)評估cho k1細(xì)胞中abc轉(zhuǎn)運(yùn)蛋白的活性。與未處理的細(xì)胞相比，用abc轉(zhuǎn)運(yùn)蛋白特異性抑制劑（圖中陰影部分）處理可誘導(dǎo)染料在細(xì)胞內(nèi)的滯留（圖中實(shí)線部分）。平均熒光強(qiáng)度（mfi）的差異表明了對應(yīng)蛋白的活性，以多藥耐藥性活性因子值（maf）表征多藥耐藥性。結(jié)果表明efluxx-id®染料對抑制劑具有良好的特異性，而calcein am染料（常用的mdr檢測探針）無法檢測bcrp活性。
部分產(chǎn)品文獻(xiàn)引用
1.overexpression of p-glycoprotein and mrp-1 are pharmacogenomic biomarkers to determine steroid resistant phenotype in childhood idiopathic nephrotic syndrome: p. narayan, et al.; pharmacogenomics j. 21, 566 (2021)
2.targeting poor proteasomal function with radioiodine eliminates ct26 colon cancer stem cells resistant to bortezomib therapy: j. h. lee, et al.; sci. rep. 10, 14308 (2020)
3.class iii β-tubulin overexpression induces chemoresistance to eribulin in a leiomyosarcoma cell line: k. yahiro, et al.; anal. cell. pathol. (amst.) 2018, 8987568 (2018), application(s): flow cytometry. mdr1 activity in human leiomysarcoma cell line sk-lms-1
4.soluble uric acid increases pdzk1 and abcg2 expression in human intestinal cell lines via the tlr4-nlrp3 inflammasome and pi3k/akt signaling pathway: m. chen, et al.; arthritis res. ther. 20, 20 (2018)
5.abcb1 and abcg2 drug transporters are differentially expressed in non-small cell lung cancers (nsclc) and expression is modified by cisplatin treatment via altered wnt signaling: m. vesel, et al.; respir. res. 18, 52 (2017)
6. expression and activity of multidrug resistance proteins in mature endothelial cells and their precursors: a challenging correlation: a. krawczenko, et al.; plos one 12, e0172371 (2017), application(s): comparison of different mdr detection methods in endothelial non-cancerous cells
7. identification of volasertib-resistant mechanism and evaluation of combination effects with volasertib and other agents on acute myeloid leukemia: y. adachi, et al.; oncotarget 8, 78452 (2017), application(s): flow cytometry; acute myeloid leukemia
8. isomahanine induces endoplasmic reticulum stress and simultaneously triggers p38 mapk-mediated apoptosis and autophagy in multidrug-resistant human oral squamous cell carcinoma cells: t. utaipan, et al.; oncol. rep. 37, 1243 (2017), application(s): flow cytometry; oral squamous cell carcinoma (oscc)
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