Highthroughput approaches to studying mechanisms of trp. Molecular devices introduces flipr calcium 6 assay kits. Fluo8 calcium flux assay kit ab112129 is a nowash, fluorescencebased assay for detecting intracellular calcium mobilization in cells from mammals and other species. R8191 flipr calcium 6 bulk kit flipr calcium 6 assay kits, molecular devices each. Intracellular calcium measurements, flipr calcium assay. Flipr calcium 3 assay kit molecular devices online store. A phenolsulphonephthalein dye in the kit forms a very stable blue colored complex specifically with free calcium.
Development and optimization of flipr high throughput. This new kit utilizes a novel masking dye technology which is exclusively licensed to molecular devices from bayer ag patent nos. The cytosolic calcium was also determined by using the flipr calcium 6 assay kit molecular devices, usa to examine the cells treated with. Enhance your calcium screens with flipr calcium 6 assay kits. Explore this application note to get basic protocol flipr calcium assay kit which is used to determine a doseresponse of carbachol in cho m1wt3 cells transfected with the muscarinic m1receptor. Flipr calcium 4 assay kit, bulk from molecular devices llc. Flipr assays for gpcr and ion channel targets assay. In the calcium flux assay protocol, cells are preloaded with fluo8 which can cross the cell membrane. Built as the most comprehensive calcium portfolio, the flipr calcium assay kits measure changes in intracellular calcium during drug discovery and research. Functional evaluation of potassium ion channels in a cell is of critical importance in the drug discovery process, particularly when it involves cardiac safety. This chapter describes the application of a protocol for the study and characterization of statedependent blockers of voltagegated calcium channels vgcc on the flipr tetra. Calcium assay kit and the bd calcium assay kit with the flipr calcium 3 assay kit using human native rhabdomyosarcoma cells expressing the urotensinii receptor ut.
Calcium signaling with flipr calcium 6 and 6qf assay kits. Flipr calcium 3 assay kit have an increased signal intensity as compared to fluo4 and fluo3 wash protocols. Flipr calcium assay kits from molecular devices employ sensitive calcium indicators and proprietary masking dyes to enable researchers to conduct highly sensitive fluorescent screens of gprotein coupled receptors gpcrs, ion channels, and other calcium sensitive targets. Although calcium assays are robust and easily amenable to hts, there are some important limitations. Configuration description part number explorer kit 10 vials of component a 1 bo. The bd calcium assay kit gives the best performance in the true nowash mode, in which both agonist and antagonist activity are easily quantified. Flipr calcium 3 assay kit demonstrates superior results from chemokine and small peptide targets. Activation of this pathway is usually measured by detection of intracellular calcium using fluorescent calcium indicator dyes and fluorescence plate readers flipr. Highthroughput approaches to studying mechanisms of trp channel activation trp channels. We have evaluated the flipr calcium 3 assay kit calcium 3, a new nowash fluorescence calcium indicator dye reagent, for the measurement of agoniststimulated calcium signaling in cells expressing the serotonin 2c 5ht2c, metabotropic glutamate receptor 5 mglur5 and the vasopressin 2 v2 gproteincoupled receptors. The calcium 6qf assay kit provides a quenchfree option for sensitive targets or multiplexing applications.
Calcium assay kit product information catalog number. Jinfang liao, chunmei wei, xing han and zhenjun diwu. Calcium indicator, 10 vials, lyophilized 10x signal enhancer, 100 ml description the bdtm calcium assay kit allows homogeneous measurement of intracellular calcium changes caused by activation of gprotein coupled receptors or calcium channels. Bavachin protects human aortic smooth muscle cells. Flipr calcium assay kits provide the most comprehensive method for measuring changes of intracellular calcium for drug discovery and basic research and act as. Identification of statedependent blockers for voltage. Screen quest calbryte520 probenecidfree and washfree. They reduce welltowell variability, improving assay quality zfactor and reliability cv % of highthroughput screens. They deliver preoptimized, homogeneous, fluorescencebased formulations to expedite assay development and screening of gpcr and ion channel targets. Molecular devices flipr calcium 6 assay kit manufacturer. With this kit, calcium assays on flipr become a mix and read procedure in which cells are incubated with the kit reagents for up to one hour and transferred directly to flipr for evaluation. If this larger book has the info and charts contained in calcium kit, then the larger, more complete book is a better place to start, contentwise. Evaluation of flipr calcium assays for screening gpcr and calcium channel targets.
Flipr calcium assay kit r8033 from molecular devices llc. The calcium 6 assay kit significantly reduces fluorescence background with a onestep protocol. Quest fluo8 am provides an optimized assay method for monitoring gpcrs and calcium channels. Lastly, the new assay data correlated better with agonistinduced calcium responses in flipr r2 0. The flipr calcium 6 assay kit fluorophore is designed to be better retained within the cell.
Configuration description part number explorer kit. Calcium flux was measured with the flexstation 3 microplate reader using the flex read mode. Calcium mobilization was measured using the flipr assay kit, after activation of the receptors by an appropriate agonist cck8s, etorphine, and dopamine, respectively traditionally, gpcrs were believed to function as monomers a single receptor binding one g protein, but, over the past decade, a new consensus has emerged in light of growing. Calcium 3 yielded equivalent 5ht2c or superior mglur5 and v2. The flipr calcium 4 assay was developed with a new inert quencher that significantly 0 reduces ligand interference, increases signal quality, minimizes the addition artifact, and works well with and without media removal before dye loading of the cells. Bioassay systems calcium assay kit is designed to measure calcium directly in biological samples without any pretreatment. Intracellular calcium measurements, flipr calcium assay kit molecular devices author. A proprietary, watersoluble probenecid is supplied with the fluo4 direct calcium assay kits, which is commonly used to inhibit this transport and reduce the baseline.
The flipr calcium assay kit from molecular devices corporation was developed and optimized for flipr to provide a fast, simple and reliable fluorescence based assay for detecting changes in intracellular calcium. Flipr calcium 5 assay kit molecular devices online store. Fluoforte calcium assay is visibly brighter enzo life sciences. The system provides a reliable and flexible high throughput screening solution for identifying early leads in the drug discovery process. Fluo4 direct calcium assay kits 2 another source of potential fluorescence outside the cells is extrusion of the indicator out of the cell by organic anion transporters. Enhance your calcium screens with flipr calcium 6 assay. Homogeneous solution for gpcr assays with flipr calcium 5. Flipr calcium 5 assay kit a novel fluorophore that amplifies the assay signal window while leveraging the superior masking technology used in the flipr calcium 4 assay kit to. R8191 flipr calcium 6 bulk kit flipr calcium 6 assay. App highlightcalcium signalling with flipr calcium 6. Now in its fourth generation, the flipr tetra is an.
Screen quest calbryte 520 probenecidfree and washfree calcium assay kit, flipr calcium 3 assay kit, flipr calcium 4 assay kit, flipr calcium 5 assay kit, invitrogen fluo4 direct, fluo4nw, no wash calcium assay kit, bd calcium assay kit. Homogeneous solution for gpcr assays with flipr calcium 5 assay kit. This htscompatible duplexed assay increases your screening capabilities, removes challenges of running multiple. Calcium releaseactivated channel crac assays on the. Figure 5 illustrates that the flipr calcium 6 assay kit with no probenecid produces a smaller signal, however the assay. Hela cells kept in continual culture were plated at. This new kit utilizes a novel masking dye technology which is exclusively licensed to molecular devices from. The flipr tetra high throughput cellular screening system flipr tetra system is a kinetic platebased, cellular assay screening system that utilizes calcium fluorescent dyes to measure calcium flux and detect cardiomyocyte beating activity 1, 2. The results were then compared to those obtained with a direct. Devices employ sensitive calcium indicators and proprietary masking dyes to enable researchers to conduct highly. Ip3ip1 assays assay guidance manual ncbi bookshelf. The flipr is a high throughput imaging plate reader that has contributed. Development and optimization of flipr high throughput calcium.
Compared to bd ratiometric calcium kit and traditional fura2 wash assay, the fura 2 qbt calcium kit provides the largest signal window and strongest z factors at ec 80 and ic 80. The flipr calcium 5 assay kit dye is also expelled from the cell and no response is seen without probenecid. Flipr tetra highthroughput cellular screening molecular. Flipr penta highthroughput cellular screening system. Chapter 12 highthroughput approaches to studying mechanisms of trp channel activation. A flipr assay for evaluating agonists and antagonists of. To address the challenging requirements of an expanding number of gpcrs and ligands, molecular devices introduces the flipr calcium 4 assay kit. Universal mixandread protocol accelerates assay workflow and increases throughput. Evaluation of flipr calcium 3 assay kita new nowash.