{"product_id":"revvity-62kbd01pea","title":"Revvity, 62KBD01PEA, HTRF Kinase-GST Discovery Kit","description":"\u003cp\u003eOverview\u003cbr\u003e\n​The best known kinase inhibitor, Gleevec (Imatinib), binds a non-activated form of Abl. Such inhibitors are therefore more difficult to detect using activity based assays, so it is worthwhile to set up kinase binding assays for your kinase of interest in order to avoid missing out on such compounds.\u003cbr\u003e\nHTRF® kinase binding assays are easily set up, using a mix and measure detection format in the absence of ATP.\u003cbr\u003e\nDirect displacement of the fluorescent-inhibitor from the ATP binding pocket is measured, enabling the affinity of your inhibitor to be determined by performing dose response curves.\u003cbr\u003e\nPurified kinase preparation is less critical compared to enzymatic assays, since measurements can be performed on kinases which have little or no activity.\u003cbr\u003e\nHow it works\u003cbr\u003e\nKinase-GST discovery kit assay principle\u003cbr\u003e\nThe binding of the tracers is detected in a sandwich assay format using a specific Anti GST antibody labeled with Europium Cryptate (donor), which binds to GST-tagged Kinase, and a red fluorescent tracer labelled with d2 (acceptor). The HTRF ratio (665\/620) will increase upon the addition of more of the tracer, and will saturate depending on the dissociation constant (Kd) of the tracer to the GST-tagged kinase. The Kinase-GST binding Discovery Kit helps determine which tracer might be best suited to setting up your binding assay . The tracer with the best assay properties (depending on the Kd and assay window generated) will be chosen to perform competitive binding assays.\u003c\/p\u003e\n\n\n  \u003cbr\u003e\n        \n\n\n\t\u003cbr\u003e\n  \u003cbr\u003e\n        \u003cbr\u003e\n  \n\n      \u003cbr\u003e\n  \u003cbr\u003e\n  \u003cbr\u003e\n    \u003cbr\u003e\n      \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n      \u003cbr\u003e\n    \n\n    \u003cbr\u003e\n      \n\n      \u003cbr\u003e\n        \n\n      \u003cbr\u003e\n    \n\n  \n\n\n\n\n\u003cp\u003e \u003cbr\u003e\nKinase-GST discovery kit assay protocol\u003cbr\u003e\nSaturation binding experiments with the three tracers (i.e. Staurosporine-Red, Dasatinib-Red, and Sunitinib-Red) can be run on 96- or 384-well plates (20 µL final volume). First, a dilution series ranging between 0 and 1 µM of tracer in the Kinase Binding Buffer is prepared in a 96-well non-binding plate. Next, 5 µL of Kinase Binding Buffer are dispensed into the final 96- or 384-well plate. Then 5 µL of GST tagged-Kinase are added, followed by 5 µL of Anti-GST Eu-cryptate. Finally, 5 µL of the red tracer solution are added. The HTRF ratio is measured after 1 H of incubation. Application-Biochemical Enzymatic Assay\u003cbr\u003e\nBrand-HTRF\u003cbr\u003e\nDetection Modality-HTRF\u003cbr\u003e\nProduct Group-Kit\u003cbr\u003e\nShipping Conditions-Shipped in Dry Ice\u003cbr\u003e\nTarget Class-Kinases\u003cbr\u003e\nTarget Species-Human\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Metabolism\/Diabetes Neuroscience Oncology \u0026amp; Inflammation\u003cbr\u003e\nUnit Size-1 unit\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775285907625,"sku":"62KBD01PEA","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-62kbd01pea","provider":"Iright","version":"1.0","type":"link"}