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Author:
Date:
2021-02-05
[Abstract] Signal transduction is the process by which molecular signals are transmitted from the cell surface to its interior, resulting in functional changes inside the cell. B cell receptor (BCR) signaling is of crucial importance for B cells, as it regulates their differentiation, selection, survival, cellular activation and proliferation. Upon BCR engagement by antigen several protein kinases, lipases and linker molecules become phosphorylated. Phosphoflow cytometry (phosphoflow) is a flow cytometry-based method allowing for analysis of protein phosphorylation in single cells. Due to recent advances in methodology and antibody availability – together with the relatively easy quantification of phosphorylation – phosphoflow is increasingly and more commonly used, compared to classical ...
[摘要] [背景] 信号转导是分子信号从细胞表面传递到细胞内部的过程，导致细胞内部功能发生变化。B细胞受体（BCR）信号对于B细胞至关重要，因为它调节了它们的分化，选择，存活，细胞活化和增殖。抗原与BCR结合后，一些蛋白激酶，脂肪酶和接头分子会被磷酸化。磷酸流式细胞仪（phosphoflow）是一种基于流式细胞仪的方法，可以分析单个细胞中的蛋白质磷酸化。由于方法学和抗体的可获得性的最新进展，再加上相对容易的磷酸化定量，与传统的蛋白质印迹分析相比，越来越多地使用磷酸流。但是，建立一种适用于所有感兴趣目标的方法可能具有挑战性。在这里，我们提出了逐步的磷流方案，允许对信号分子的磷酸化状态进行评估，并进行广泛的染色，以鉴定各种人和鼠B细胞亚群，如先前在Rip等人的原始论文。，（2020 ）。在原始论文中对磷流靶标的描述旁边，我们提供了在BCR信号传导中起关键作用的其他靶标的指导。循序渐进的磷酸流操作流程是用户友好的操作，可对人和鼠B细胞亚群中各种BCR信号分子的磷酸化进行灵敏检测。
[背景] ...
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