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Reactivity: The CD45 monoclonal antibody is directed against the CD45- antigen, defined T200 or Leucocyte Common Antigen. The antibody reacts with all cells of the haemopoietic lineage, not with cells of other lineages.
The CD14 monoclonal antibody is directed against the CD14- antigen, which is expressed on human monocytes and macrophages. The antibody reacts with human monocytes and macrophages; weak reactions may occur with neutrophils.
Specificity: The CD45 antibody recognizes 180, 195, 205, 220, kD MW components of the leucocyte common antigen complex to be found on lymphocytes, monocytes, granulocytes, thymocytes and malignant T and B cells. No reactivity has been observed with primary or metastatic carcinoma cells. Plasma cells or myeloma cells may have weak expression or be negative for this antigen.
The CD14 antibody recognizes 55kD MW monocyte surface antigen identified by monoclonal antibodies belonging to the CD14 cluster and found on peripheral blood monocytes and weakly on granulocytes. Monocytic cells, interfollicular dendritic cells and dendritic reticulum cells are stained in sections of lymphoid tissue sections.
Clinical applications: Abnormal numbers of cells expressing the target antigen or cells showing aberrant expression levels of the antigen can be expected in some disease states. This may result in an altered pattern of staining.
It is important to understand the normal expression pattern of an antigen and its relationship to the expression of other relevant antigens in order to perform appropriate analysis. Nonlymphocyte components of LWB that may contaminate the lymphocyte analysis gate include monocytes, granulocytes (neutrophils, eosinophils, basophils), and debris (residual erythrocytes, erythrocyte ghosts, and platelets).
The lymphocyte analysis gate should be set to include the largest possible number of lymphocytes in the sample. This is done to avoid biasing the data by selectively excluding lymphocyte subsets exhibiting slightly different light-scatter patterns than the majority of lymphocytes.