Abstract：
Detection of phosphorylated proteins in tissue sections using immunohistochemistry (IHC) is a challenging task. The absence of tissue staining may be caused by either a lack of protein expression or a lack of protein activation via its phosphorylation. To address this problem, we employed Integrated Co-detection Workflow (ICW) protocol to analyze lung cancer tissue sections by combining in situ hybridization (ISH) with IHC. The target protein of interest was epidermal growth factor receptor (EGFR, also known as ErbB1 and HER1) which is the founding member of the ErbB family of receptor tyrosine kinases. Using phospho-specific antibodies specific for a phosphorylated site Y1173 of EGFR molecule allowed us to analyze IHC and ISH staining at a single cell level in lung cancer tissue. We have observed both a co-localization of IHC with ISH signals and ISH-positive cells lacking IHC labeling for phosphorylated EGFR. ICW appears to be a very powerful spatial biology technique for accurate localization of cancer cells with phosphorylated/activated and non-phosphorylated/nonactivated proteins.
摘要：
使用免疫组织化学(IHC)检测组织切片中的磷酸化蛋白是具有挑战性的任务。组织染色的缺失可能是由于缺乏蛋白质表达或缺乏通过其磷酸化的蛋白质活化引起的。为了解决这个问题,我们采用集成共检测工作流(ICW)方案,通过结合原位杂交(ISH)和IHC分析肺癌组织切片.目的靶蛋白是表皮生长因子受体（EGFR，也称为ErbB1和HER1），是受体酪氨酸激酶ErbB家族的创始成员。使用对EGFR分子的磷酸化位点Y1173具有特异性的磷酸特异性抗体允许我们在肺癌组织中以单细胞水平分析IHC和ISH染色。我们已经观察到IHC与ISH信号的共定位和缺乏磷酸化EGFR的IHC标记的ISH阳性细胞。ICW似乎是一种非常强大的空间生物学技术，可以准确定位具有磷酸化/活化和非磷酸化/非活化蛋白的癌细胞。
