Journal
BIO-PROTOCOL
Volume 11, Issue 22, Pages -Publisher
BIO-PROTOCOL
DOI: 10.21769/BioProtoc.4230
Keywords
CD45; Diabetes; Kidney; KIM-1; IHC; OVE
Categories
Funding
- JDRF [1-2005-88, 3-2005-932]
- NIH
- NIDDK [DK072032]
- JDRF
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In this study, a reliable immunohistochemistry (IHC) protocol for CD45 staining in paraffin-embedded mouse kidney sections was shared. The protocol demonstrated its utility in pathology research by significantly increasing the number of CD45-positive cells in the kidneys, indicating the ability of the CD45 antibody to recognize bone marrow cells and macrophages.
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.
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