Binch Abbie, Snuggs Joseph, Le Maitre Christine L
Biomolecular Sciences Research Centre Sheffield Hallam University Sheffield UK.
JOR Spine. 2020 Jun 11;3(3):e1098. doi: 10.1002/jsp2.1098. eCollection 2020 Sep.
Immunohistochemistry (IHC) is a useful technique for the localization and semiquantification of protein expression within tissues. Adult human intervertebral disc (IVD) tissues contain a large amount of auto-fluorescence which often makes immunofluorescence techniques inappropriate on tissue samples but can be applied to isolated cell samples. Thus, IHC remains one of, if not the most common application for protein detection within IVD tissue. Immunostaining localizes antigen expression through specific epitope-antibody interactions. Within the field of IVD research, IHC is commonly used on fresh frozen and paraffin embedded tissues to elucidate the expression of antigens. Here, we discuss the principles of IHC applied to formalin fixed paraffin embedded IVD tissue and supply optimized protocols for antibodies used within our group to guide research within the IVD field.
免疫组织化学(IHC)是一种用于在组织内定位和半定量蛋白质表达的有用技术。成人人类椎间盘(IVD)组织含有大量自发荧光,这常常使得免疫荧光技术不适用于组织样本,但可应用于分离的细胞样本。因此,免疫组织化学仍然是椎间盘组织内蛋白质检测的最常用应用之一(即便不是唯一最常用的)。免疫染色通过特异性表位-抗体相互作用来定位抗原表达。在椎间盘研究领域,免疫组织化学常用于新鲜冷冻和石蜡包埋组织,以阐明抗原的表达。在此,我们讨论应用于福尔马林固定石蜡包埋椎间盘组织的免疫组织化学原理,并提供我们团队使用的抗体的优化方案,以指导椎间盘领域的研究。