Stephen Gaffin writes: >Which is the "better" assay for Th1/Th2-ness? flow cytometry or >ELISA? and which are the best cytokines to measure? Better * S ease, convenience, speed, reproducibility, sensitivity There is a new method useful for evaluating Th1/Th2-ness, the MACS Cytokine Secretion Assays. The fundamental technique employed in this assay is the capture of secreted cytokine on the surface of the cell by a catch-reagent antibody (an antibody complex with bi-specificity for both the cell and cytokine secreted). A second PE labeled anti-cytokine antibody binds to the secreted, captured cytokine, and the labeled cells can be analyzed by flow cytometry, with no effect on cell viability, unlike common intracellular cytokine detection methods which permeabilize cells and destroy cell viability. In addition, the cytokine secreting cells can be selected and enriched utilizing MACS magenetic selection columns. Two MACS product exist for detecting and enriching gamma-interferon and IL-4 secreting cells. Common applications include detection and enrichment of Antigen Specific T Cells. One potential advantage of the MACS Cytokine Secretion Assays versus the very elegant tetramer assay is flexibility and ease of use. There is no need to assemble MHC-peptide complexes using the MACS technique, and also no need to ultilize specific peptide immunogen, which works well, also. Best regards, Ron Ronald P. Dudek Clinical Products Business Manager Miltenyi Biotec, Inc. 251 Auburn Ravine Rd. Suite 208 Auburn, CA 95603 Phone (800) 367-6227 Fax (530) 888-8925
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