IL-5 Antibody, anti-human, PE,

Intracellular staining of IL-5–producing human or non-human primate cellsInterleukin 5 (IL-5) is pre-dominantly secreted by CD4+ T cells. It is involved in a range of allergic reactions and mediates immune reactions against parasites. IL-5 also acts on other cell types such as B cells. | The Anti-IL-5 antibody has been designed for intracellular staining of IL-5–producing cells. Cells can be stimulated for IL-5-production, for example, by polyclonal stimulation with mitogens. For induction of IL-5 production by antigen-specific T cells, cells are restimulated with respective antigen. IL-5 can be accumulated in the cells by addition of secretion inhibitors like brefeldin A. After fixation and permeabilization of the cell sample, IL-5–producing cells can be stained intracellularly with Anti-IL-5 antibodies. Staining of surface markers allows simultaneous flow cytometric analysis of subsets and activation status of the IL-5–producing cells. Magnetically enriched cells can be stained intracellularly for IL-5 production directly on the MACS® Column. This procedure ensures higher sensitivity of detection and minimizes loss of cells during washing procedures. The protocol is useful for cytokine analysis of rare cells, for example, CD4+ T cells in HIV patients, or other cell sources than peripheral blood mononuclear cells (PBMCs), e.g., bronchoalveolar lavages or synovial fluids.