通过流式细胞术测定生物素化抗体的亲和力。
Affinity determination of biotinylated antibodies by flow cytometry.
作者信息
Even-Desrumeaux Klervi, Chames Patrick
机构信息
INSERM U624, Marseille, France.
出版信息
Methods Mol Biol. 2012;907:443-9. doi: 10.1007/978-1-61779-974-7_25.
Affinity determination is a crucial step of an antibody characterization. Here, we describe a method for antibody affinity determination by flow cytometry, relying on the unique affinity of biotin for streptavidin for easy and efficient antibody labeling. Several labeling approaches are described and discussed in this chapter, including chemical and enzymatic (in vivo and in vitro) biotinylation. Finally, a procedure for K (D) determination by flow cytometry is precisely described.
亲和力测定是抗体表征的关键步骤。在此，我们描述一种通过流式细胞术测定抗体亲和力的方法，该方法依赖于生物素与链霉亲和素的独特亲和力，以实现简便高效的抗体标记。本章描述并讨论了几种标记方法，包括化学和酶促（体内和体外）生物素化。最后，详细描述了通过流式细胞术测定解离常数（KD）的步骤。
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