Rhod-4 No Wash Calcium Assay Kit (ab112157) is a fluorescence-based assay for detecting intracellular calcium mobilization.
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- Advanced science (Weinheim, Baden-Wurttemberg, Germany) 8:e21011812021Reduced Oligodendrocyte Precursor Cell Impairs Astrocytic Development in Early Life Stress.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYuxin Wang,Yixun Su,Guangdan Yu,Xiaorui Wang,Xiaoying Chen,Bin Yu,Yijun Cheng,Rui Li,Juan C Sáez,Chenju Yi,Lan Xiao,Jianqin NiuPubMed 34155833
- Investigative ophthalmology & visual science 61:442020Electrophysiologic Characterization of Developing Human Embryonic Stem Cell-Derived Photoreceptor Precursors.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRevital Schick,Nairouz Farah,Amos Markus,Alon Korngreen,Yossi MandelPubMed 32991686
- International journal of molecular sciences 21:2020Dexamethasone Treatment Increases the Intracellular Calcium Level Through in A549 Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBo-Hui Jeon et. al.PubMed 32033337
- Pain 158:931-9442017A central role for R7bp in the regulation of itch sensation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMritunjay Pandey,Jian-Hua Zhang,Santosh K Mishra,Poorni R Adikaram,Benjamin Harris,John F Kahler,Anna Loshakov,Roxanne Sholevar,Allison Genis,Claire Kittock,Juraj Kabat,Sundar Ganesan,Richard R Neubig,Mark A Hoon,William F SimondsPubMed 28134655
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Cell-based
- Reactive species
- Mammals
- Assay time
- 1h
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Target data
Alternative names
Ca++, Ca2+
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Rhod-4 No Wash Calcium Assay Kit (ab112157) is a fluorescence-based assay for detecting intracellular calcium mobilization.
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Cell-based
- 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 No Wash Calcium Assay Kit (ab112157) 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. ab112157 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) or 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 - No Wash (ab112157)
Carbachol Dose Response was measured in HEK293 cells with ab112157 and Rhod-2. HEK293 cells were seeded overnight at 40,000 cells/100 µL/well in a black wall/clear bottom 96-well plate. The cells were incubated with 100 µL of dye-loading solution using ab112157, 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 0.6 µM, and that for Rhod-2 is about 0.7 µM.
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