TNFα (mouse) AlphaLISA Detection Kit, 500 Assay Points

Formats:

• 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:

• No-wash steps, no separation steps
• ELISA alternative technology
• Sensitive detection
• Broad sample compatibility
• Small sample volume
• Results in less than 3 hours
• Half the time of an ELISA assay

In the mouse, Tumor Necrosis Factor alpha (TNFa) is primarily produced as a homotrimeric 235 amino acid membrane- bound protein. The soluble mature homotrimeric form of 156 amino acids is then released by the metalloprotease TNFa converting enzyme. In humans, TNFa is produced by many cell types like macrophages, monocytes, neutrophils, T cells, and NK cells. It causes cytolysis and cytostasis of many tumor cell lines in vitro. Within hours of injection, TNFa leads to the destruction of small blood vessels within malignant tumors. Although TNFa inhibits the growth of endothelial cells in vitro, it is a potent promoter of angiogenesis in vivo. In contrast to chemotherapeutic drugs, TNFa specifically attacks malignant cells. Furthermore, TNFa is associated with autoimmune disorders, and antibodies directed against TNFa have proven useful.

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.