Development and optimization of cytokine ELISAs using commercial antibody pairs
Development and optimization of cytokine ELISAs using commercial antibody pairs
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DOI：
10.1016/s0022-1759(01)00419-7
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发表时间：
2001-09-01
影响因子：
2.2
通讯作者：
Remick, DG
中科院分区：
文献类型：
--
作者：
Nemzek, JA;Siddiqui, J;Remick, DG
血浆和其他液体中细胞因子的测量通常需要使用酶联免疫吸附测定（ELISA）。在研究环境中，一个有价值的分析是在最短的时间内以最低的成本产生可靠的结果。为了实现这一目标，已经概述了使用商业抗体开发用于细胞因子的夹心ELISA的方案。这些ELISA开发指南包括选择抗体浓度、选择适当的缓冲液、减少血浆干扰和评估孵育期的最佳长度。此外，一个快速的IL-6 ELISA的协议。该ELISA通过提高所用抗体的浓度和提高孵育温度，允许在减少的时间量内测量IL-6。通过遵循所提出的指南，可以开发出具有成本效益的细胞因子ELISA，其产生低背景，检测宽范围的浓度，并且适用于研究环境。(C)2001 Elsevier Science B. V.保留所有权利。
The measurement of cytokines in plasma and other fluids often requires the use of an enzyme-linked immunosorbant assay (ELISA). In the research environment, a valuable assay is one that yields reliable results in the shortest amount of time for the least cost. To achieve this goal, a protocol has been outlined to develop sandwich ELISAs for cytokines using commercial antibodies. These guidelines for ELISA development include selecting antibody concentrations, choosing an appropriate buffer, reducing plasma interference and evaluating the optimal length for incubation periods. In addition, the protocol for a rapid IL-6 ELISA is presented. This ELISA allows measurement of IL-6 in a reduced amount of time by raising the concentration of antibodies used and increasing the temperature for incubation. By following the guidelines presented, cost-effective, cytokine ELISAs can be developed that yield low background, detect a wide range of concentrations, and are suitable for use in the research setting. (C) 2001 Elsevier Science B.V. All rights reserved.