Fluo-8 Medium Removal Calcium Assay kit (ab112128)

Overview

  • Product nameFluo-8 Medium Removal Calcium Assay kitSee all Calcium kits ...
  • Sample type
    Adherent cells, Suspension cells
  • Assay typeCell-based
  • Species reactivity
    Reacts with: Human
  • Product overview

    Abcam's Fluo-8 Calcium Assay Kits provide homogeneous fluorescence-based assays for detecting intracellular calcium mobilization. Cells expressing a GPCR of interest that signals through calcium are pre-loaded with Fluo-8 which can cross the cell membrane. Once inside the cell, the lipophilic blocking groups of Fluo-8 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 Fluo-8. The characteristics of its long wavelength, high sensitivity, and >100 times fluorescence enhancement make Fluo-8 an ideal indicator for the measurement of cellular calcium.
    ab112128 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 ab112128 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.

Fluo-8 Medium Removal Calcium Assay kit images

  • Carbachol Dose Response was measured in HEK293 cells with ab112128 and a Fluo-4 assay kit. HEK293 cells were seeded overnight at 40,000 cells/100 µL/well in a black wall/clear bottom 96-well plate. The growth medium was removed, and the cells were incubated with 100 µL of dye-loading solution using ab112128 or the Fluo-4 kit for 1 hour at room temperature. Carbachol (25 µL/well) was added to achieve the final indicated concentrations. The EC50 of ab112128 is about 1.2 µM.

Protocols

References for Fluo-8 Medium Removal Calcium Assay kit (ab112128)

ab112128 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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The lab confirmed that the formula is correct.

I hope this is of help.

Thank you for contacting us and your interest in our products.

The Calcium Quantification Assay kit (Fluorometric), ab112115, is intended for use with solution based samples. For your experiment, it may be better to have a look at the Fluo-8 M...

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Thank you for contacting us.

The reagents in ab112128 are sufficient for one 96-well plate, not five.

Regarding the Greiner Bio-one plate, catalogue number is 655090, you could use it, however it is much better to use Poly-D-Lysin...

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Yes, that plate will be suitable.
Please let me know if you have any other questions or concerns.

Following up my previous reply, I have the following information from the developer of the assay:

The calcium assay in cells is a transient response, so you will need a bottom-read fluorescent plate reader with a built-in pipetting system su...

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Thank you for your inquiry.

No, the plate is not included with the kit. If you need a recommendation for a plate, could you please tell me if you are working with adherent cells or non-adherent cells?

Thank you for your inquiry.

We do not know of any publications that used ab112128 at this point in time. As soon as we receive news about a publication we will up-date the datasheet.

I would like to reassure your customer that we gu...

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