Abcam Rhod-4 Calcium Assay kit - Medium Removal No Wash (ab112156) is a fluorescence-based assay for detecting intracellular calcium mobilization.
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Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Quantitative
- Reactive species
- Mammals
- Assay time
- 1h
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Target data
Alternative names
Ca++, Ca2+
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Abcam Rhod-4 Calcium Assay kit - Medium Removal No Wash (ab112156) is a fluorescence-based assay for detecting intracellular calcium mobilization.
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Quantitative
- Reactive species
- Mammals
- Assay time
- 1h
- Assay Platform
- Microplate reader
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
Abcam Rhod-4 Calcium Assay kit - Medium Removal No Wash (ab112156) is a fluorescence-based assay for detecting intracellular calcium mobilization. Cells expressing a GPCR of interest that signals through calcium are pre-loaded with Rhod-4 which can cross the cell membrane. Once inside the cell, the lipophilic blocking groups of Rhod-4 are cleaved by an esterase, resulting in a negatively charged fluorescent dye that stays inside the cell. Its fluorescence is greatly enhanced upon binding to calcium. When cells are stimulated with agonists, the receptor signals the release of intracellular calcium, which significantly increases the fluorescence of Rhod-4. The characteristics of its long wavelength, high sensitivity, and >250 times fluorescence enhancement make Rhod-4 the brightest red calcium indicator available in the marker, and it is an ideal tool for the measurement of cellular calcium through HTS screening. Compared to Fluo-8, Rhod-4 is more photostable, making its fluorescence imaging more robust. ab112156 provides an optimized assay method for monitoring the G-protein-coupled receptors and calcium channels using HTS instrumentation. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format. Visit our for tips and troubleshooting.
This product is intended to be used for monitoring calcium fluctuations in vivo in live cells using the following HTS imaging plate readers: FLIPR™, FDSS, BMG NOVOstar™, FLexStation, ViewLux, IN Cell Analyzer or Arrayscan.
If you would like to quantify calcium concentration in vitro using cell extracts, we recommend using Calcium Detection Kit (Colorimetric) (Calcium Assay Kit (Colorimetric) ab102505) Calcium Quantification Assay Kit (Calcium Assay kit (Fluorometric) ab112115), as they provide stock standards for comparison.
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Functional Studies - Rhod-4 Calcium Assay kit - Medium Removal No Wash (ab112156)
Carbachol Dose Response measured in HEK293 cells using Rhod-4 Medium Removal No Wash Calcium Assay kit and Rhod-2.
HEK293 cells were incubated with 100 µL of the dye-loading solution using ab112156, or 100 µL of Rhod-2 solution (5 µM) for 1 hour at room temperature. Carbachol (25 µL/well) was added to achieve the final indicated concentrations. The EC50 of Rhod-4 is about 0.8 µM, and that for Rhod-2 about 0.7 µM.
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