Hellström Cecilia, Dodig-Crnković Tea, Hong Mun-Gwan, Schwenk Jochen M, Nilsson Peter, Sjöberg Ronald
Affinity Proteomics, School of Biotechnology, SciLifeLab, KTH-Royal Institute of Technology, Box 1031, SE-17121, Stockholm, Sweden.
Affinity Proteomics, SciLifeLab, School of Biotechnology, KTH - Royal Institute of Technology, Box 1031, 17121, Solna, Sweden.
Methods Mol Biol. 2017;1619:229-238. doi: 10.1007/978-1-4939-7057-5_18.
In-depth exploration and characterization of human serum and plasma proteomes is an attractive strategy for the identification of potential prognostic or diagnostic biomarkers. The possibility of analyzing larger numbers of samples in a high-throughput fashion has markedly increased with affinity-based microarrays, thus providing higher statistical power to these biomarker studies. Here, we describe a protocol for high-density serum and plasma reverse phase protein arrays (RPPAs). We demonstrate how a biobank of 12,392 samples was immobilized and analyzed on a single microarray slide, allowing high-quality profiling of abundant target proteins across all samples in one assay.
深入探索和表征人血清和血浆蛋白质组是识别潜在预后或诊断生物标志物的一种有吸引力的策略。基于亲和的微阵列以高通量方式分析大量样本的可能性显著增加,从而为这些生物标志物研究提供了更高的统计效力。在此,我们描述了一种用于高密度血清和血浆反相蛋白质阵列(RPPA)的方案。我们展示了如何将一个包含12392个样本的生物样本库固定在一张微阵列载玻片上并进行分析,从而在一次检测中对所有样本中的丰富靶蛋白进行高质量分析。