TNFα (human) AlphaLISA Biotin-Free Detection Kit, 5,000 Assay Points

Formats:

• Our HV (100 assay point) kits allow you to run 100 wells in 96-well format, using a 100 µL reaction volume (10 µL of sample).
• Our 500 assay point kit allows you to run 500 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).
• Our 5,000 assay point kit allows you to run 5,000 wells in 96-well or 384-well format, using a 50 µL reaction volume (5 µL of sample).

Features and Benefits:

• No-wash steps, no separation steps
• Compatible with high biotin samples, including RPMI and high-biotin tissue extracts
• ELISA alternative technology
• Sensitive detection
• Broad sample compatibility
• Results in less than 3 hours

Tumor Necrosis Factor Alpha is a multifunctional proinflammatory cytokine synthesized mainly by nucleated blood cells as a 233 aa type II transmembrane protein which is cleaved by ADAM17 between aa76-77 to form a soluble homotrimeric complex. TNFα plays a role in lipid metabolism, coagulation and endothelial function and has been associated with cancer, infection and inflammation (including inflammatory bowel disease), ischemia/reperfusion injury and heart failure, and insulin resistance. This kit is used when the analyte is in a matrix containing high levels of biotin, such as can be found in cell culture media such as RPMI, or milk.

AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA biotin-free assay, a DIG-labeled anti-analyte antibody binds to the anti-DIG-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.