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Phospho-PDGFRβ (Tyr751) Cellular Assay Kit Homogeneous cell-based assay for PDGF receptor β involved in angiogenesis and cancer  Cisbio's cell-based homogeneous HTRF® phospho-PDGFRβ assay (Tyr751) enables rapid, quantitative detection of the modulation of the platelet-derived growth factor receptor beta, phosphorylated on Tyrosine 751. PDGFRbeta signals and crosstalks via both the PI3K/AKT and the Ras/Raf MAPK pathways. This cell surface tyrosine kinase receptor ensures the binding of particular growth factors, which trigger the MAPK pathway leading to cell proliferation, cell differentiation and angiogenesis. When activated, PDGFRbeta forms a dimer and becomes phosphorylated, which makes the phospho-PDGFRβ assay a valuable tool in oncology research. This simple add-and-read assay is ideal for screening small molecule compounds or biologics directly in cells. The phospho-PDGFRβ ready-to-use kit contains all the reagents you need and offers enhanced convenience over other immunoassay technologies.    Features Cell-based sandwich immunoassay Validated on the basal endogenous level of PDGFRbeta Compatible with human, mouse and rat cellular models Truly simple homogeneous protocol with no wash steps More convenient and faster than ELISA or WB   Applications Cellular pharmacological quantification of phospho-PDGFR-β Tyr751   Functional screening of PDGF receptor modulators From pathway exploration to preclinical drug discovery phases Angiogenesis, cancer, stroke, fibrosis and infectious diseases     Assay Principle HTRF® - the homogeneous cell-based sandwich immunoassay Cisbio's phospho-PDGFR-beta assay is based on a TR-FRET sandwich immunoassay format comprising two specific anti-PDGFR-beta antibodies, one labeled with a cryptate as donor and the other with d2 as acceptor. The phospho-PDGFR-beta antibodies bind the phosphorylated residue, and the proximity of donor and acceptor will then lead to a fluorescent TR-FRET signal. The signal intensity is proportional to the substrate phosphorylation. The protocol is optimized for a 384-well plate format, but can easily be further miniaturized or upscaled. Only low sample volumes are needed. The detection reagents may be pre-mixed and added in a single dispensing step for direct detection. No washing is needed at any step.   The phospho-PDGFR-beta assay kit can be run with frozen cell lysates or fresh cells in culture. After cell lysis, phospho-PDGFR-beta can be quantitatively detected using the HTRF phospho-PDGFR-beta kit reagents and most TR-FRET multimode plate readers.     Two-plate assay protocol For added flexibility, the assay can be run under a two-plate assay protocol, where cells are plated and treated in a 96-well culture plate. For the detection of phosphorylated PDGFR-beta, lysates are subsequently transferred to a 384 small volume assay plate where the HTRF reagents are added. This also enables the monitoring of cell viability and confluence in an appropriate cell culture plate.     

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