一种用于检测人Bcl10蛋白的高灵敏度微孔板酶联免疫吸附测定法的开发、评估及应用。
Development, evaluation, and application of a highly sensitive microtiter plate ELISA for human Bcl10 protein.
作者信息
Bhattacharyya Sumit, Pant Nitika, Dudeja Pradeep K, Tobacman Joanne K
机构信息
Department of Medicine, University of Illinois at Chicago and Jesse Brown VA Medical Center, Chicago, IL, 60612 USA.
出版信息
J Immunoassay Immunochem. 2007;28(3):173-88. doi: 10.1080/15321810701454573.
Bcl10 (B-cell CLL/lymphoma 10) is a 233 amino acid CARD (caspase recruitment domain)-containing cellular protein, increasingly recognized as a mediator of NFkappaB activation in non-immune, as well as immune cells. Due to the importance of Bcl10 in diverse cell types, we developed a solid-phase, enzyme-linked immunosorbent (ELISA) assay to precisely measure Bcl10 in small volume cell lysates, using recombinant Bcl10 to standardize the assay. Standard curve measures Bcl10 from 0.25 ng/mL to 16 ng/mL, with very low intra- and inter-assay variation. Sample dilution and exogenous Bcl10 recovery experiments, comparisons with Western blot, and linear response to increasing doses of known Bcl10 activators confirm the specificity and precision of the ELISA.
Bcl10（B细胞慢性淋巴细胞白血病/淋巴瘤10）是一种含有233个氨基酸的含半胱天冬酶招募结构域（CARD）的细胞蛋白，在非免疫细胞和免疫细胞中越来越被认为是NFκB激活的介质。由于Bcl10在多种细胞类型中的重要性，我们开发了一种固相酶联免疫吸附（ELISA）测定法，使用重组Bcl10对该测定法进行标准化，以精确测量小体积细胞裂解物中的Bcl10。标准曲线可测量0.25 ng/mL至16 ng/mL的Bcl10，测定内和测定间的变异非常低。样品稀释和外源性Bcl10回收实验、与蛋白质印迹法的比较以及对已知Bcl10激活剂剂量增加的线性反应证实了ELISA的特异性和精确性。
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