Description
Introduction:
The KRIBIOLISA ELISA kits are used for assessing the specific biomarker in samples analytes which may be serum and cell culture supernatant as validated with the kit. The kit employs a blocking ELISA technique which engages the blocking pathway to estimate the neutralizing antibodies. Tenecteplase is a tissue plasminogen activator (tPA) developed from modifications of natural human tPA complementary DNA (cDNA). It is a 527 amino acid with a substitution of threonine 103 with asparagine and substitution of asparagine 117 with glutamine within the kringle 1 domain, and a tetra-alanine substitution at amino acids 296-299 in the protease domain. A neutralizing antibody (NAb) is an antibody that is responsible for defending cells from pathogens and are produced naturally by the body as part of its immune response. Their production is triggered by both infections and vaccinations against infections. In an immunogenetic context it will bind to a drug and neutralize its therapeutic effect.
Intended Use:
The KRIBIOLISA Neutralizing Antibodies to Tenecteplase (TNKase) ELISA kit is used as an analytical tool for the qualitative detection of neutralizing antibodies against Tenecteplase in serum or plasma.
Principle:
The method employs sandwich ELISA technique. The protein-protein interaction between HRP-Tenecteplase and Anti-TPA can be blocked by Tenecteplase. The neutralizing antibodies against Tenecteplase binds to Tenecteplase and thus a competitive inhibition is created. Samples and controls are pipetted in a blank microtitre plate and incubated with neutralizing antibody to Tenecteplase (NAb) and Tenecteplase. This complex of bound and unbound Tenecteplase is then incubated with HRP conjugated Tenecteplase protein. The unbound Tenecteplase will bind to the HRP conjugated Tenecteplase. The bound Nab-Tenecteplase will not bind to the HRP conjugated Tenecteplase. This complex solution of bound antibodies to Tenecteplase and unbound Tenecteplase is then pipetted into Anti-TPA coated microplate. After washing, the substrate solution (TMB) is added to the microwells. Post incubation, color develops proportionate to the amount of bound Tenecteplase. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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