Research assay kit FSP-0102-P4-1

tuberculosisTBimmunoassay

Paves your way to functional analysis of a specific research question. Intended use This FluoroSpot Path kit is intended for the enumeration of cells secreting human IFN-γ and IL-2 in response to TB peptides using the FluoroSpot assay. For research use only. Not for use in diagnostic procedures. Peptide pool composition The scanning pools consist of synthetic peptides of 15-mers overlapping with 11 amino acids, covering the complete proteins. The ESAT-6 scanning pool is derived from the early secretory antigenic target protein and contains 21 peptides. The CFP-10 scanning pool is derived from the culture filtrate protein and contains 23 peptides. Finally, the EspC scanning pool is derived from the ESX-1 secretion-associated protein and contains 23 peptides. Performance Human PBMC (250,000 cells/well) from an individual with latent tuberculosis were incubated overnight in the absence or presence of the ESAT-6 and CFP-10 scanning pools (Mabtech). The number of cells secreting IFN-γ and/or IL-2 was analyzed by FluoroSpot. Single cytokine secreting cells as well as double-secreting cells were determined. Spot center was used to identify spots from double-secreting cells, visualized in an image overlay of LED490, FITC (IFN-γ) and LED550,Cy3 (IL-2). Spot analysis was made with Mabtech IRIS. IFN-γ Analyte description - Interferon-γ (IFN-γ) is the only type II interferon. This proinflammatory cytokine is secreted by activated T cells and NK cells. It activates macrophages and endothelial cells and regulates immune responses by affecting APCs, T cells, and B cells. Production of IFN-γ by helper T cells and cytotoxic T cells is a hallmark of the Th1-type phenotype.