Department of Biology, School of Science, University of Duhok, Duhok, Kurdistan Regional Government, Iraq.
Merck Group, Glasgow, Scotland, UK.
Methods Mol Biol. 2020;2163:163-170. doi: 10.1007/978-1-0716-0696-4_13.
Humanized rat basophilic leukemia (RBL) reporter cell lines are increasingly used for the detection of allergen-specific IgE and other purposes, such as the detection of allergens and standardization of allergen preparations. Existing reporter systems have many strengths and advantages but can be expensive or require longer incubation times. The new NPY-mRFP reporter cell line addresses such problems, as it requires neither expensive substrates nor overnight incubation for detection of activation. The fusion of Neuropeptide Y (NPY) with monomeric Red Fluorescent Protein (mRFP) results in localization of the fluorescent protein in granules. As NPY-mRFP is preformed in granules, the reporter system activation can be assessed using fluorescence measurements after as soon as 45-60 min, as described in this chapter, without the need to add any substrates.
人源化大鼠嗜碱性白血病（RBL）报告细胞系越来越多地用于检测过敏原特异性 IgE 及其他目的，如过敏原的检测和过敏原制剂的标准化。现有的报告系统具有许多优点，但可能价格昂贵或需要更长的孵育时间。新的 NPY-mRFP 报告细胞系解决了这些问题，因为它既不需要昂贵的底物，也不需要过夜孵育即可检测激活。神经肽 Y（NPY）与单体红色荧光蛋白（mRFP）融合导致荧光蛋白在颗粒中定位。由于 NPY-mRFP 预先形成于颗粒中，因此可以在 45-60 分钟后使用荧光测量来评估报告系统的激活，如本章所述，而无需添加任何底物。
