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Wash Free Calcium Assay Kit
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.
Applications
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.
Key features and benefits
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.
Calcium influx assay principle
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.
Kit contents
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
Instrument requirement
Fluorescence plate reader with on board injectors
Learn more and purchase
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
 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
This system does not allow the amplification of DNA originated from other sources, such as cultured cells, which affect the detection result. Amplification of gene sequence with PCR using these primer pairs enhances not only the sensitivity but also the specificity of detection.
Better signal, better performance (Downloadable product manual)
HDB Wash Free Fluo-4 Calcium Assay Kits are still available for you:
 Description
 Catalog No.
 Size
 HDB WASH Free Calcium Assay Kit
 HD02-0010
 10 Plates
 HDB WASH Free Calcium Assay Kit
 HD02-0100
 100 Plates
Disclaimer: Materials from HD Biosciences are sold for research use only. Neither resale nor transfer to a third party is allowed.
Comparison Between Fluo-4 and Fluo-8 Calcium Assay Data
Figure 1. HEK293/mGluR6/Ga15 cells were seeded overnight in 100¦Ìl 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 ¡À SEM. pEC50 value was determined using GraphPad Prism 5 software.
Figure 2. HEK293/mGluR3/Ga16 and HEK293/Ga16 cell lines were seeded overnight in 100¦Ìl 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 ¡À SEM. pEC50 value was determined using GraphPad Prism 5 software. kit.
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 100¦Ìl L-Glutamate (signal) and assay buffer (background).
Figure 4. HEK293/TRPV1 cells were seeded overnight in 100¦Ìl 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 ¡À SEM. pEC50 value was determined using GraphPad Prism 5 software.
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P.R.China
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