Quantification of Protein Expression by Proximity Ligation Assay in the Nonhuman Primate in Response to Estrogen.
Quantification of Protein Expression by Proximity Ligation Assay in the Nonhuman Primate in Response to Estrogen.
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DOI：
10.1007/978-1-0716-1920-9_6
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发表时间：
2022
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在蛋白质生物学领域，基于免疫学的技术不断发展，用于检测和定量细胞和组织中的单个蛋白质水平、蛋白质-蛋白质相互作用和蛋白质修饰。邻位连接试验（PLA）是一种结合免疫学和基于PCR的方法的检测方法，其开发目的是克服其他检测方法固有的一些缺点。PLA允许在固定的组织和培养的细胞中原位非常灵敏和离散地定量测量未修饰的天然蛋白质水平和蛋白质-蛋白质相互作用/修饰复合物。我们在此描述了PLA方法及其定量雌激素对血管调节因子表达的影响的适用性，血管系统中的内皮一氧化氮合酶（eNOS）、胎盘中的血管内皮生长因子（VEGF）和非人灵长类动物胎儿肾上腺中的黑皮质素2受体（MC2R）/辅助蛋白（MRAP）。
In the field of protein biology, immunology-based techniques are continuously evolving for detection and quantification of individual protein levels, protein-protein interaction and protein modifications in cells and tissues. The proximity ligation assay (PLA), a method of detection that combines immunologic and PCR-based approaches, was developed to overcome some of the drawbacks that are inherent with other detection methods. The PLA allows for very sensitive and discretely quantifiable measures of unmodified, native protein levels and protein-protein interaction/modification complexes in situ in both fixed tissues and cultured cells. We describe herein the PLA method and its applicability to quantify the effects of estrogen on expression of angioregulatory factors, e.g., endothelial nitric oxide synthase (eNOS) in the vasculature, vascular endothelial growth factor (VEGF) in the placenta, and melanocortin 2 receptor (MC2R)/accessory protein (MRAP) in the fetal adrenal of the nonhuman primate.