{"product_id":"revvity-62kb01rede","title":"Revvity, 62KB01REDE, HTRF Staurosporine Red Ligand, 20 nmoles","description":"\u003cp\u003eOverview\u003cbr\u003e\nThis compound is derived from Staurosporine, which is a natural alkoid present in Streptomyces Staurosporeus. It is a prototypical ATP-competitive Kinase inhibitor, and has been shown to be highly promiscuous and therefore very useful to address multiple Kinases in the HTRF Kinase Binding assay platform. The ‘d2’ fluorescent dye used absorbs with its maximum near 650 nm and fluoresces near 670 nm. It is an acceptor molecule implemented in HTRF assays. \u003cbr\u003e\nHTRF assays offer many advantages over other technologies:\u003c\/p\u003e\n\n\u003cp\u003eHomogeneous add-and-read format\u003cbr\u003e\nNo wash steps\u003cbr\u003e\nLow background\u003cbr\u003e\nStraightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates\u003cbr\u003e\nStable signal, providing flexibility in time of readout or size of assays\u003cbr\u003e\nHow it works\u003cbr\u003e\nKd determination assay principal\u003cbr\u003e\nThe binding of Staurosporine-Red is detected in a sandwich assay format using the Anti Tag labeled with Europium Cryptate (donor) which binds 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 of the tracer, and will saturate depending on the dissociation constant (Kd) of the tracer to the tagged kinase.To know whether Staurosporine-Red is the tracer  best suited to your tagged kinase of interest, we advise you to first try the Kinase Binding Discovery kits. \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\nKd determination assay protocol\u003cbr\u003e\nSaturation binding experiments of  Staurosporine-Red can be run in 96- or 384-well plates (20 µL final volume).\u003cbr\u003e\nFirst, a dilution series ranging between 0 and 1  µM of Staurosporine-Red in the Kinase Binding Buffer is prepared in a 96- well non-binding plate.\u003cbr\u003e\nNext, 5 µL of Kinase Binding Buffer are dispensed into the final 96- or 384-well plate. Then 5 µL of tagged-Kinase are added, followed by 5 µL of Anti-tag Eu-cryptate. Finally, 5 µL of the red tracer solution are added. \u003cbr\u003e\nThe HTRF ratio is measured after 1H of incubation.\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\nIC50\/Ki determination assay principle\u003cbr\u003e\nThe binding of Staurosporine-Red is detected in a sandwich assay format using a specific Anti-GST, 6HIS antibody or Streptavidin labeled with Europium Cryptate (donor), which binds to the tagged Kinase, and a red fluorescent Staurosporine labelled with d2 (acceptor).The detection principle is based on HTRF® technology.  The HTRF ratio (665\/620) will increase with the addition of more Staurosporine-Red, and will saturate depending on the dissociation constant (Kd ) of Staurosporine-Red to the tagged kinase. When an inhibitor of the kinase is added, Staurosporine-Red will be displaced and the HTRF signal will disappear, depending on the dose.\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\nIC50\/Ki determination assay protocol\u003cbr\u003e\n​Pharmacological evaluations of inhibitors of interest can be run in 96- or 384-well plates.\u003cbr\u003e\nFirst a dilution series of the inhibitor ranging between 40 µM and 0.23 nM is prepared, and  5 µL of each concentration are dispensed into the plate. Then 5 µL of tagged-Kinase are added, followed by 5 µL of anti-tag Eu-cryptate. Finally, 5 µL of Staurosporine-Red solution are added, prepared at 4x the final concentration.  The HTRF ratio is measured after 1H 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-Fluorescent Reagent\u003cbr\u003e\nShipping Conditions-Shipped in Dry Ice\u003cbr\u003e\nTarget Class-Kinases\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Metabolism\/Diabetes Neuroscience Oncology \u0026amp; Inflammation\u003cbr\u003e\nUnit Size-20 nmol\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775317364905,"sku":"62KB01REDE","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-62kb01rede","provider":"Iright","version":"1.0","type":"link"}