CD45 Immunohistochemistry in Mouse Kidney

Area of Science:

• Immunohistochemistry
• Cell Biology
• Pathology

Background:

• CD45 is a crucial pan-leukocyte marker used to assess inflammatory cell infiltration and its link to tissue damage.
• Standardized protocols are essential for accurate CD45 staining in research settings.

Purpose of the Study:

• To establish and validate a reliable immunohistochemistry (IHC) protocol for CD45 staining in paraffin-embedded mouse kidney sections.
• To demonstrate the utility of this protocol in identifying inflammatory cells in kidney injury models.

Main Methods:

• Utilized a rat anti-CD45 antibody as the primary antibody.
• Employed a biotin-conjugated goat anti-rat IgG secondary antibody.
• Implemented a horseradish peroxidase (HRP)-linked avidin/biotin detection system with 3,3'-Diaminobenzidine (DAB) for signal visualization.

Main Results:

• The protocol successfully identified hematolymphoid cells in bone marrow and monocyte/macrophages in liver and lung tissues.
• Significantly increased CD45-positive cells were observed in the kidneys of diabetic mice, validating the protocol's utility.
• Double staining revealed CD45-positive cells accumulating around injured tubular cells and overlapping with other inflammatory cells like macrophages.

Conclusions:

• This validated IHC protocol provides a robust method for CD45 staining in mouse kidney tissue.
• The protocol is effective for quantifying inflammatory cell infiltration in various tissues and disease models, particularly in kidney injury research.
• The findings highlight the importance of CD45 staining in understanding inflammatory responses in pathological conditions.