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(gè)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(dòng)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(huì)減弱。EFLUXX-ID®多藥耐藥性分析試劑盒是目前少有可同時(shí)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

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(shí)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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