{"product_id":"revvity-62kbbrdf","title":"Revvity, 62KBBRDF, HTRF Kinase Binding Buffer, 200 ml","description":"\u003cp\u003eOverview\u003cbr\u003e\n​The Kinase Binding Buffer is used to perform saturation binding (Kd-determination) and\/or competitive binding assays within the HTRF® Kinase Binding platform only.\u003cbr\u003e\nHow it works\u003cbr\u003e\nKd determination assay principal\u003cbr\u003e\nThe binding of a fluorescent tracer is detected in a sandwich assay format using either a specific Anti-GST, 6HIS antibody or Streptavidin labeled with Europium Cryptate (donor), which bind to the tagged Kinase, and a red fluorescent tracer labelled with d2 (acceptor). The detection principle is based on HTRF® technology. The HTRF ratio (665\/620) will increase upon the addition of more tracer, and will saturate depending on the dissociation constant (Kd ) of the tracer to the tagged kinase. When an inhibitor of the kinase is added, the tracer will be displaced and the HTRF signal will disappear, depending on the dose.\u003c\/p\u003e\n\n\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 \u003c\/p\u003e\n\n\n\n\n\u003cp\u003eIC50\/Ki determination assay protocol\u003cbr\u003e\n​Pharmacological evaluation of inhibitors of interest can be run in 96- or 384-well plates.\u003cbr\u003e\nFirst, a dilution series of inhibitor ranging between 40 µM and 0.23 nM is prepared, and 5 µL of each concentration are dispensed into the plate. Next, 5 µL of tagged-Kinase are added, followed by 5 µL of anti-tag Eu-cryptate. Finally, 5 µL of tracer solution are added, prepared at 4x the final concentration. The HTRF ratio is measured after 1 H of incubation.\u003cbr\u003e\nAnalyses of the data give typical dose response curves ranging between 10 µM and 56 pM, enabling an evaluation of the IC50\/Ki values for the inhibitor of interest. Application-Biochemical Enzymatic Assay\u003cbr\u003e\nBrand-HTRF\u003cbr\u003e\nDetection Modality-HTRF\u003cbr\u003e\nProduct Group-Buffer\u003cbr\u003e\nShipping Conditions-Shipped Ambient\u003cbr\u003e\nTarget Class-Kinases\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Metabolism\/Diabetes Neuroscience Oncology \u0026amp; Inflammation\u003cbr\u003e\nUnit Size-200 mL\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775285809321,"sku":"62KBBRDF","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-62kbbrdf","provider":"Iright","version":"1.0","type":"link"}