| Measurement of Transferrin- and Non-transferrin-bound Iron Uptake by Mouse Tissues
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Author:
Date:
2016-09-05
[Abstract] Iron in blood plasma is bound to its transport protein transferrin, which delivers iron to most tissues. In iron overload and certain pathological conditions, the carrying capacity of transferrin can become exceeded, giving rise to non-transferrin-bound iron, which is taken up preferentially by the liver, kidney, pancreas, and heart. The measurement of tissue transferrin- and non-transferrin-bound iron (TBI and NTBI, respectively) uptake in vivo can be achieved via intravenous administration of 59Fe-labeled TBI or NTBI followed by gamma counting of various organs. Here we describe a detailed protocol for the measurement of TBI and NTBI uptake by mouse tissues.
[摘要] 血浆中的铁结合其转运蛋白转铁蛋白，其将铁递送至大多数组织。 在铁过载和某些病理状况下，转铁蛋白的携带能力可能超过，导致非转铁蛋白结合的铁，其优先被肝脏，肾脏，胰腺和心脏摄取。 分别测量组织转铁蛋白和非转铁蛋白结合的铁（分别为TBI和NTBI）在体内的摄取可以通过静脉内施用59 Fe标记的TBI或 NTBI，然后是各种器官的γ计数。 在这里我们描述了测量小鼠组织的TBI和NTBI摄取的详细协议。
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| XTT Assay of Antifungal Activity
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Author:
Date:
2015-08-05
[Abstract] XTT assay is a colorimetric method that uses the tetrazolium dye, 2,3-bis-(2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanilide (XTT) to quantify cell-mediated damage to fungi. Actively respiring fungal cells convert the water-soluble XTT to a water-soluble, orange colored formazan product (Meshulam et al., 1995). Here, we describe the protocol that measures the ability of plasmacytoid dendritic cells (pDCs) to exert antifungal activity. This approach was first established with human polymorphonuclear cells (PMN) by Meshulam et al. (1995) and then adapted to pDC by Ramirez-Ortiz et al. (2011) and Loures et al. (2015). It can be modified for use with other effector cells and to test compounds for antifungal activity.
[摘要] XTT测定是使用四唑鎓染料，2,3-双 - （2-甲氧基-4-硝基-5-磺酰基） - （2H） - 四唑鎓-5-甲酰苯胺（XTT）定量细胞介导的损伤的比色法 到真菌。 主动呼吸真菌细胞将水溶性XTT转化为水溶性橙色甲an产物（Meshulam等人，1995）。 在这里，我们描述测量浆细胞样树突状细胞（pDCs）发挥抗真菌活性的能力的协议。 该方法首先由Meshulam等人（1995）的人多形核细胞（PMN）建立，然后由Ramirez-Ortiz等人（2011）修改为pDC， Loures 等人（2015）。 其可以被修饰以与其他效应细胞一起使用并且测试化合物的抗真菌活性。
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