Trk B (Phospho-Tyr515)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA kit from Assay Genie designed for the detection of phosphorylation of Trk B at tyrosine 515. The assay uses a colorimetric detection method measured at 450 nm and is reactive with Human, Mouse and Rat samples. It is presented as a convenient, lysate-free, high-throughput format suited to detecting Trk B protein phosphorylation and expression profiles in cultured adherent cells, with a dynamic range above 5000 cells.

The assay follows an indirect ELISA format in which Trk B (Phospho-Tyr515) is captured by target-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. Substrate addition triggers a measurable colorimetric reaction catalysed by the conjugated HRP. Two normalisation methods are supplied: a monoclonal antibody specific for human GAPDH acts as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining method can be used to determine cell density and adjust for plating differences.

Background: TrkB (gene symbol NTRK2, Gene ID 4915, UniProt Q16620) is a receptor tyrosine kinase of the Trk family that serves as a receptor for brain-derived neurotrophic factor (BDNF) and neurotrophin-3, -4 and -5, and is involved in the development and maintenance of the nervous system. Phosphorylation at tyrosine 515 is described as critical for activating downstream signalling cascades that regulate neuronal survival, growth and differentiation, with dysregulation of TRK-B signalling implicated in various neurological disorders.