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Human Interleukin 10 (IL10) is a homodimer composed of two subunits of 18 kDa each. It is produced by various T cell populations, monocytes, macrophages, and different cell types in the liver when stimulated by endogenous or exogenous factors such as stress, exotoxins, tumor necrosis factor-a, and catecholamines. IL10 inhibits interferon-? synthesis in Th1 cell clones, monocytes and macrophages. IL10 also inhibits antigen presentation to T cells and IL12 production by monocytes. It also impairs the proliferation and cytokine synthesis of CD4+ T cells, without having a direct inhibitory effect on CD8+ T cells. On the other hand, IL10 has a stimulatory effect on B cells, prevents apoptosis and enhances proliferation and differentiation of plasma cells, and inhibits the release of various chemokines by neutrophils. In general, the main biological function of IL10 is to limit and terminate the inflammatory responses, block proinflammatory cytokine secretion and regulate the differentiation and proliferation of several immune cells. IL10 activity is mediated by the heteromeric IL10 receptor (IL-10R), and signals through the tyrosine kinases Jak1 and Tyk2, and STATs.
AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
|Assay Target Class||Cytokine|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5,000 assay points|
The introduction of enzyme-linked immunosorbent assays (ELISAs) in the early 1970’s offered researchers a non-radiometric immunoassay platform without compromising sensitivity. Over the last 50 years scientists have made huge strides in disease research and drug discovery and a demand for greater assay throughput and sensitivity has evolved. In response, more robust immunoassays have been developed to address some of the limitations of the standard, colorimetric ELISA.
Find out about the most common limitations of traditional ELISAs and how different ELISA alternative technologies address these limitations.
Biomarker levels were measured directly in CulturPlates-96 and-384 in a simple, fast, all-in-one-well AlphaLISA assay format. The elimination of transfer and wash steps simplifies cellular assays, reduces variability and significantly reduces hands-on time and costs associated with consumables. Integral plasma membrane (EGFR) and secreted solubleproteins (TNFa, IL1ß, IL6, IL8) were successfully determined, on adherent or suspension cells, using the standard AlphaLISA Immunoassay buffer.
Enzyme-linked Immunosorbent Assay is the most widely Kits adopted method for detection and quantification of cytokines and other biomarkers. This traditional technology offers good,selectivity, sensitivity and assay versatility; however, it has certain disadvantages such as limited dynamic range and low throughput due to the numerous wash steps.