• Product name
    Fluo-8 Calcium Flux Assay Kit - No Wash
    See all Calcium kits
  • Detection method
  • Sample type
    Adherent cells, Suspension cells
  • Assay type
  • Assay time
    1h 00m
  • Assay duration
    One step assay
  • Product overview

    Fluo-8 Calcium Assay Kit - No Wash (ab112129) is a fluorescence-based assay for detecting intracellular calcium mobilization in cells from mammals and other species.

    Cells 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.

    If intracellular calcium levels increase, the fluorescence of Fluo-8 is significantly increased. The characteristics of its long wavelength, high sensitivity, and >100 times fluorescence enhancement make Fluo-8 the brightest green calcium indicator available, and it is an ideal tool for the measurement of cellular calcium through HTS screening.

    ab112129 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 FAQs page for tips and troubleshooting.

  • Notes

    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.

    Readers with an in built pipetter are essential for measuring calcium response accurately due to transient property of the calcium signal. Depending on dose of the agonist, the peak response is few seconds to 20 seconds, so one will need to add the compound by instrument and read the fluorescent signal simultaneously every second for ~1-2 min.

    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).

  • Platform



  • Carbachol Dose Response was measured in HEK293 cells with ab112129 and a Fluo-4 calcium assay kit. 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 the ab112129 or the Fluo-4 kit for 1 hour at room temperature. Carbachol (50 µL/well) was added to achieve the final concentrations indicated.



This product has been referenced in:
  • Dyer A  et al. Oncolytic Group B Adenovirus Enadenotucirev Mediates Non-apoptotic Cell Death with Membrane Disruption and Release of Inflammatory Mediators. Mol Ther Oncolytics 4:18-30 (2017). Read more (PubMed: 28345021) »
  • Xie Y  et al. Upregulation of RGS2: a new mechanism for pirfenidone amelioration of pulmonary fibrosis. Respir Res 17:103 (2016). Read more (PubMed: 27549302) »

See all 2 Publications for this product

Customer reviews and Q&As

Thank you for your enquiry by telephone yesterday and for your patience while I contacted the laboratories.

They have confirmed that this is correct, the Pluronic F127 Plus component is red in color in the tube.

The fluorescence that ...

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Thank you for your message and I am sorry to hear you have had difficulty obtaining results from ab112129 Fluo-8 No Wash Calcium Assay kit

The only fluorescence microplate readers recommended for ab112129 Fluo-8 No Wash Calcium Assay kit a...

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Use black wall clear bottom 96 well cell culture treated plates.

Yes, the kit contains Probenecid. No additional Probenecid is needed for the kit.

The Ca response is a transient response (a couple of the seconds to 10-20 seconds), it is almost impossible to pipette the compounds into the plate, and read the response. Yes, you do use the multichannel pipette to add the dye into the plate, and let ...

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Yes, the kit can be used for all the mammalian cells - the lab did confirm this.

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 contacting us.

The kit uses our water soluble and heat stable probeneic to prevent dyes from leaking out of the cells.

I hope this information is helpful to you. Please do not hesitate to contact us if you...

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Thank you very much for your email.

I realize that there was a typos in my last email. The kD value I should have written is 320nm. However I also contacted laboratory after receiving your reply - they have informed me that the kD value is 389...

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Thank you for contacting us.

It seems that you have asked similar question last year in October and we provided the 230nm as dissociation constant. The 230 nm Kd value is available to us from our collaborator who developed this product for us....

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1-10 of 17 Abreviews or Q&A


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