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用于测量小鼠体内细胞因子产生的体内细胞因子捕获试验。

The in vivo cytokine capture assay for measurement of cytokine production in the mouse.

作者信息

Finkelman Fred, Morris Suzanne, Orekhova Tatyana, Sehy David

机构信息

University of Cincinnati College of Medicine and Cincinnati Veterans Administration Medical Center, Cincinnati, Ohio, USA.

出版信息

Curr Protoc Immunol. 2003 Aug;Chapter 6:Unit 6.28. doi: 10.1002/0471142735.im0628s54.

Abstract

Because most cytokines are utilized, catabolized, or excreted shortly after they are produced, it has been difficult to directly measure in vivo cytokine production. Consequently, it has been necessary to infer in vivo cytokine secretion levels from the results of ex vivo assays of cytokine secretion, assays that measure tissue levels of cytokine mRNA, or assays that stain tissues for cytokine protein levels. Results of these assays provide important and useful information, but do not necessarily reflect in vivo cytokine secretion. To better determine in vivo cytokine production, the in vivo cytokine capture assay (IVCCA) was developed. IVCCA facilitates measurement of cytokines in serum by increasing their in vivo half-lives. This increases the sensitivity of measurement of in vivo cytokine production 30- to 1,000-fold. The first protocol described in this unit is for luminescence-based ELISA, while the second is for an absorbance-based method.

摘要

由于大多数细胞因子在产生后不久就被利用、分解代谢或排泄,因此很难直接测量体内细胞因子的产生。因此,有必要从细胞因子分泌的体外检测结果、测量细胞因子mRNA组织水平的检测或对细胞因子蛋白水平进行组织染色的检测结果中推断体内细胞因子的分泌水平。这些检测结果提供了重要且有用的信息,但不一定反映体内细胞因子的分泌情况。为了更好地确定体内细胞因子的产生,开发了体内细胞因子捕获检测法(IVCCA)。IVCCA通过延长细胞因子在体内的半衰期来促进血清中细胞因子的测量。这将体内细胞因子产生测量的灵敏度提高了30至1000倍。本单元描述的第一个方案是基于发光的ELISA,而第二个方案是基于吸光度的方法。

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