| Good news for researchers around the world: HD Biosciences successfully launched Wash Free Fluo-8 Calcium Assay kits to the market. The Wash free Fluo-8 Calcium Assay kits provide much higher signal intensity and assay robustness compared with other commercially available calcium influx assay kits. Fluo-8 AM is currently the brightest calcium indicator, more than 2 fold brighter than Fluo-4 AM, and 4 times brighter than Fluo-3 AM. With this kit, dye loading can be performed at room temperature, and calcium assays on fluorometric plate reader become a mix-and-read procedure in which cells are incubated with the kit reagents for one hour and transferred directly to plate reader for evaluation. There are no intermediate wash-steps involved. The Fluo-8 Calcium Assay Kit from HDB provides a fast, simple and reliable fluorescence-based assay for detecting changes in intracellular calcium. |
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| Applications
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| The kit provides a homogeneous assay for calcium flux. It is specially designed to work for a broad range of targets, including GPCRs and ion channels. The kits have been successfully used with both adherent and suspension cells. |
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| Key features and benefits |
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 Increased Signal Intensity: with the brightest calcium indicator Fluo-8.
Rapid dye loading: Dye loading can be performed at room temperature.
Convenient and Robust: No wash steps involved
Ideal for all calcium influx assays: Compatible with many cell lines and targets without ligand or target interference.
Adaptable to high throughput screening: 96-well or 384-well plate format compatible.
Cost effective: Very competitive price. |
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| Calcium influx assay principle |
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| This kit provides a homogenous assay format for detection of calcium mobilization. Cells expressing a GPCR of interest or of channels that signal through calcium are pre-loaded with a calcium-sensitive dye and are then treated with compound. Upon stimulation, the receptor signals the release of intracellular calcium, which results in a large fluorescence increase of the calcium sensitive dye upon binding to calcium. The signal generated in the assay is read using fluorescent plate readers that possess an injector for the addition of agonist and is capable of a rapid read of the resulting change in fluorescence. The assay has been designed to accommodate both 96 well and 384 well formats, which are commonly used in high throughput cell-based functional screening. |
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| Kit contents |
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HDB Wash Free Fluo-8 Calcium Assay Component A
HDB Wash Free Fluo-8 Calcium Assay Component B
HDB Wash Free Fluo-8 Calcium Assay Component C
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| Instrument requirement |
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| Fluorescence plate reader with on board injectors |
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| Learn more and purchase |
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Kits are available in two sizes providing sufficient material for either 10 plates (96 or 384) or 100 plates. Additionally, for higher volume customers, custom packaged materials can also be provided upon request. Please contact: operation@hdbiosciences.com or call: (+86)-21-58558330 |
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| Description |
Catalog No. |
Size |
| HDB WASH Free Fluo-8 Calcium Assay Kit |
HD03-0010 |
10 Plates |
| HDB WASH Free Fluo-8 Calcium Assay Kit |
HD03-0100 |
100 Plates |
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| Better signal, better performance (Downloadable product manual) |
HDB Wash Free Fluo-4 Calcium Assay Kits are still available for you:
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| Description |
Catalog No. |
Size |
| HDB WASH Free Calcium Assay Kit |
HD02-0010 |
10 Plates |
| HDB WASH Free Calcium Assay Kit |
HD02-0100 |
100 Plates |
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Disclaimer: Materials from HD Biosciences are sold for research use only. Neither resale nor transfer to a third party is allowed. |
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Comparison Between Fluo-4 and Fluo-8 Calcium Assay Data |
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Figure 1. HEK293/mGluR6/Ga15 cells were seeded overnight in 100μl on a 96-well Matrigel coated plate. Assays with Wash Free HDB Fluo-4 and Fluo-8 calcium assay kits were performed according to the protocols described in the kit manuals. Data `points represent means ± SEM. pEC50 value was determined using GraphPad Prism 5 software. |
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| Figure 2. HEK293/mGluR3/Ga16 and HEK293/Ga16 cell lines were seeded overnight in 100μl on a 96-well Matrigel coated plate. Assays with Wash Free HDB Fluo-4 and Fluo-8 NW calcium assay kits were performed according to the protocols described in the kit manuals. Data points represent means ± SEM. pEC50 value was determined using GraphPad Prism 5 software. kit. |
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Fig 3. HDB WASH Free Calcium Assay Kit shows consistent pharmacology for known agonist and antagonist. |
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| Figure 3. Z’ factor determination in 96-well format for HEK293/mGluR3/Ga16 cells were performed with Wash Free HDB Fluo-4 and Fluo-8 NW calcium assay kits. HEK293/mGluR3/Ga16 cell line was stimulated with 100μl L-Glutamate (signal) and assay buffer (background). |
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| Figure 4. HEK293/TRPV1 cells were seeded overnight in 100μl on a 96-well Matrigel coated plate. Assays with Wash Free HDB Fluo-4 and Fluo-8 NW calcium assay kits were performed according to the protocols described in the kit manuals. Data points represent means ± SEM. pEC50 value was determined using GraphPad Prism 5 software. |
