{"product_id":"revvity-pt8haloneo","title":"Revvity, PT8HALONEO, Tag-lite pT8-HaloTag (Neomycin) Plasmid, 10 µg","description":"\u003cp\u003eOverview\u003cbr\u003e\nOver the past few years, SNAP-Tag technology combined with TR-FRET has paved the road to the development of many non-radioactive, no-wash binding assays. The method is based on transfecting cells using plasmids encoding a SNAP-Tag and subsequently labeling them with Terbium. PT8HALONEO is a backbone containing the gene for the HALO tag, a promoter a neomycin resistance gene and MCS (Multiple Cloning Site).\u003cbr\u003e\nTag-lite assays offer many features and applications over traditional receptor\/ligand binding assays:\u003c\/p\u003e\n\n\u003cp\u003eNon-radioactive\u003cbr\u003e\nHomogeneous and filtration-free\u003cbr\u003e\nReady-to-use kits and reagents for binding assays\u003cbr\u003e\nDoes not alter the receptor pharmacology\u003cbr\u003e\nPeer-reviewed, validated technology\u003cbr\u003e\nThorough kinetic studies with true Kd and Ki values, association (Kon) and dissociation (Koff) rate constants\u003cbr\u003e\nScreening and profiling of biologics and large molecules\u003cbr\u003e\nHow it works\u003cbr\u003e\nStep 1 - Plasmid creation\u003cbr\u003e\nUsing standard cloning techniques, insert the GPCR gene of interest into the empty plasmid. Remember that when you are designing a plasmid, the GPCR gene should be kept in the frame.\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\u003cp\u003eStep 2 - Plasmid transfection\u003cbr\u003e\nUse standard transfection techniques (see transient transfection protocol) to transiently express the SNAP-GPCR of interest in your cell line.\u003c\/p\u003e\n\n\n\u003cp\u003eStep 3 - Receptor labeling\u003cbr\u003e\nSNAP-tag®, is a small fusion tag that covalently interacts with specific substrates. SNAP-tag allows specific and covalent labeling of any protein of interest. Cells are provided unlabeled and need to be labeled with Lumi4-Terbium prior to running a binding assay. Labeling reagents are available from the Revvity catalog in 4 different sizes. For more details see the labeling procedure.\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\u003eStep 4 - Understand the assay principle\u003cbr\u003e\nRunning a receptor binding assay using Tag-lite is as easy as it can get. Simply dispense 10 µL of labeled cells into each well, followed by 5 µL of labeled ligand and 5 µL of the compound you wish to test. Like all HTRF assays, Tag-lite assays do not require any washing steps. A diagram of the procedure to be followed is given on the right.\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\u003cp\u003eStep 5 - Saturation binding (KD)\u003cbr\u003e\nA saturation binding assay measures total and non-specific binding for increasing concentrations of ligand under equilibrium conditions. To perform the assay, the fluorescent ligand is titrated into a solution containing a fixed amount of labeled cells and then incubated to equilibrium. The HTRF ratio obtained from this titration is the total binding.\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  \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\u003cp\u003eStep 6 - Competitive binding (KI)\u003cbr\u003e\nA competitive binding assay is performed to measure the dissociation constant, Ki. To perform the assay, the compound is titrated into a solution containing a fixed concentration of fluorescent ligand and a fixed amount of cells. Application-Receptor-Ligand Binding\u003cbr\u003e\nBrand-Tag-lite\u003cbr\u003e\nDetection Modality-HTRF\u003cbr\u003e\nProduct Group-Plasmids\u003cbr\u003e\nShipping Conditions-Shipped in Dry Ice\u003cbr\u003e\nTarget Class-GPCR\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Cardiovascular Infectious Diseases Metabolism\/Diabetes NASH\/Fibrosis Neuroscience Oncology \u0026amp; Inflammation Rare Diseases\u003cbr\u003e\nUnit Size-10 mg\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775301931177,"sku":"PT8HALONEO","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-pt8haloneo","provider":"Iright","version":"1.0","type":"link"}