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AlphaLISA technology allows the detection of molecules of interest in buffer, cell culture media, serum and plasma in a highly sensitive, quantitative, reproducible and user-friendly mode. In an AlphaLISA assay, a Biotinylated Anti-Analyte Antibody binds to the Streptavidin-coated Alpha 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 provokes 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.
TNFR1 is a transmembrane receptor of TNF-α and TNF-β. The soluble form (sTNFR1) is released by the cell in inflammatory response. sTNFR1 interacts with free TNF-α to inhibit inflammation. Many neurological disorders are associated with high levels of serum TNFR1 such as, schizophrenia, bipolar, and dementia. Mutations of the gene which encodes TNFR1 may lead to tumor necrosis factor receptor associated periodic syndrome (TRAPS) and/or multiple sclerosis.
|Assay Target Class||Protein|
|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.
Biomarkers provide insight into the presence and progression of disease states. Tumor necrosis factor 1 (TNF1) is a transmembrane protein that can be released by extracellular proteolytic cleavage to function as cytokines. Increased levels of soluble TNFR1 is an indicator for disease states such as infection, type 2 diabetes, and can be used as a predictive marker for progression to advanced chronic kidney disease or end-stage renal disease.
Typically, wash-based ELISAs are used to quantify increased levels of soluble TNFR1 from complex patient samples. While sensitive, ELISAs often require large amounts of samples and can be very labor intensive. AlphaLISA® is a homogenous bead-based assay. This technical note demonstrates how to quantify levels of TNFR1 in human urine samples by comparing normal patient donors to type 2 diabetics.
See how AlphaLISA technology provides a rapid, homogeneous, bead-based assay format for the quantification of an analyte in complex matrices.
Tumor Necrosis Factors (TNFs) are cytokines that are the primary modifiers of inflammatory and immune response. Researchers have shown that a soluble form of TNFR1 (sTNFR1) is a truncated version of the receptor produced by the disintegration and extracellular release of membranous protein on the cell surface (ectodomain shedding). sTNFR1 is found in healthy and diseased patients alike, however increased sTNFR1 levels are an indicator for disease states such as inflammation, infection, and asthma.
Here we present a way to measure TNFR1 using homogeneous bead-based AlphaLISA assay. The human TNFR1 AlphaLISA® detection kit was designed for the quantitative determination of soluble TNFR1 in serum and cell culture media. This technical note further demonstrates the functionality of the kit by detecting sTNFR1 in cell supernatant as well as TNFR1 on the cell membrane.