Overview

  • Product nameRhod-4 No Wash Calcium Assay kitSee all Calcium kits ...
  • Sample type
    Adherent cells, Suspension cells
  • Assay typeQuantitative
  • Assay time
    1h 00m
  • Species reactivity
    Reacts with: Human
  • Product overview

    Abcam’s Calcium Assay Kits provide a homogeneous fluorescence-based assay for detecting the intracellular calcium mobilization. Cells expressing a GPCR of interest that signals through calcium are pre-loaded with our proprietary Rhod-4 which can cross cell membranes. Rhod-4 is the brightest red calcium indicator available for HTS screening. Once inside the cell, the lipophilic blocking groups of Rhod-4 are cleaved by non-specific cell esterases, resulting in a negatively charged fluorescent dye that stays inside the cells. Its fluorescence is greatly enhanced upon binding to calcium. When cells are stimulated with screening compounds, the receptor signals the release of intracellular calcium, which greatly increase the fluorescence of Rhod-4.

    ab112157 provides an optimized assay method for monitoring G-protein-coupled receptors and calcium channels. The kit comes with all the essential components and an optimized protocol. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation.

    Visit our FAQs page for tips and troubleshooting.

  • Notes

    This product is intended to use for monitoring calcium fluctuations in vivo in live cells. If you would like to quantify calcium concentration in vitro using cell extracts, we recommend using Calcium Detection Kit (Colorimetric) (ab102505) or Calcium Quantification Assay Kit (ab112115), as they provide stock standards for comparison.

  • Tested applicationsFunctional Studies more details

Properties

  • RelevanceCalcium is essential for all living organisms, where Ca2+ sequestration and release into and out of the cytoplasm functions as a signal for many cellular processes. 99% of calcium is found in bones and teeth with the remaining 1% found in the blood and soft tissue. Serum calcium levels are tightly controlled (8.4-11.4 mg/dL) and any variation outside this range can have serious effects. Calcium plays a role in mediating the constriction and relaxation of blood vessels, nerve impulse transmission, muscle contraction, and hormone secretion. Calcium ion channels control the migration of calcium ions across cell membranes, permitting the activation and inhibition of a wide variety of enzymes. Causes of low calcium levels include chronic kidney failure, vitamin D deficiency, and low blood magnesium levels that can occur in severe alcoholism.
  • Alternative names
    • Ca++
    • Ca2+

Applications

Our Abpromise guarantee covers the use of ab112157 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
Functional Studies Use at an assay dependent concentration.

Rhod-4 No Wash Calcium Assay kit images

  • 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.

Protocols

References for Rhod-4 No Wash Calcium Assay kit (ab112157)

ab112157 has not yet been referenced specifically in any publications.

Product Wall

Thank you for your inquiry.

1. if the kits for measuring intracellular calcium mobilization can work with mouse cell lines, specifically MLE-12 cell line?--Yes, as long as the cells contains calcium channels and Gq receptors

2. Also, ...

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Thank you for getting back to me.

Yes, the signal would be expected to be enhanced in the presence of zinc.

And in reply to your following question, yes the Fluo-8 (ab112129) and Rhod-4 (ab112157) would not be suitable for your samp...

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Thank you for your inquiry.

I can confirm that you can use PBS instead of HHBS.

It is essential that20mM Hepes Buffer is added to the PBS (as it is added to the HHBS).

I wish you good luck with your experiments.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"