小鼠肾脏中的CD45免疫组织化学
CD45 Immunohistochemistry in Mouse Kidney.
作者信息
Zheng Shirong, Epstein Paul N
机构信息
Diabetes and Obesity Center, Division of Environmental Medicine, School of Medicine, University of Louisville, KY, USA.
Department of Pediatrics, School of Medicine, University of Louisville, KY, USA.
出版信息
Bio Protoc. 2021 Nov 20;11(22):e4230. doi: 10.21769/BioProtoc.4230.
CD45 is a pan-leukocyte marker, and CD45 stain is widely used to determine the extent of inflammatory cell infiltration and its association with tissue injury. In this manuscript, we share a reliable immunohistochemistry (IHC) protocol for CD45 staining in sections of paraffin-embedded mouse kidney. A rat anti-CD45 antibody was used as primary antibody, and a mouse adsorbed biotin-conjugated goat anti-rat IgG was selected as secondary antibody. A horseradish peroxidase (HRP)-linked avidin/biotin detection system was used to amplify the signal, which was detected with 3,3'-Diaminobenzidine (DAB). With this protocol, we show that the CD45 antibody recognizes cells of hematolymphoid lineage in bone marrow, as well as monocyte/macrophages in liver and lung tissue. The utility of this protocol in pathology research was indicated by dramatically increased CD45-positive (CD45) cells in the kidneys of a mouse model of diabetes. Double staining for CD45 and injury marker KIM-1 showed accumulated CD45 cells around injured tubular cells. CD45 and F4/80 macrophage staining on adjacent tissue sections revealed overlap of CD45 cells with other inflammatory cells.
CD45是一种全白细胞标志物，CD45染色被广泛用于确定炎症细胞浸润的程度及其与组织损伤的关联。在本论文中，我们分享一种用于石蜡包埋小鼠肾脏切片中CD45染色的可靠免疫组织化学（IHC）方案。使用大鼠抗CD45抗体作为一抗，并选择小鼠吸附的生物素偶联山羊抗大鼠IgG作为二抗。使用辣根过氧化物酶（HRP）标记的抗生物素蛋白/生物素检测系统来放大信号，该信号用3,3'-二氨基联苯胺（DAB）进行检测。通过该方案，我们表明CD45抗体可识别骨髓中造血淋巴系细胞，以及肝脏和肺组织中的单核细胞/巨噬细胞。糖尿病小鼠模型肾脏中CD45阳性（CD45+）细胞显著增加，表明该方案在病理学研究中的实用性。CD45与损伤标志物KIM-1的双重染色显示，损伤的肾小管细胞周围有聚集的CD45+细胞。相邻组织切片上的CD45与F4/80巨噬细胞染色显示，CD45+细胞与其他炎症细胞存在重叠。
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