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Fluo-8 No Wash Calcium Assay kit (ab112129)

Q&A (15)

Overview

  • Product nameFluo-8 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 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.

    ab112129 provides an optimized assay method for monitoring the G-protein-coupled receptors and calcium channels. 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

    ab112129 should be stored dessicated.

  • 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 ab112129 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 FuncS

    Fluo-8 No Wash Calcium Assay kit images

    • Functional Studies - Fluo-8 No Wash Calcium Assay kit (ab112129)
      Functional Studies - Fluo-8 No Wash Calcium Assay kit (ab112129)
      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.

    Protocols

    References for Fluo-8 No Wash Calcium Assay kit (ab112129)

    ab112129 has not yet been referenced specifically in any publications.

    Publishing research using ab112129 ? Please let us know so that we can cite the reference in this datasheet

    Product Wall

    Submit a Question for ab112129

    Displaying 1 - 10 of 15 results for Q&A

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    What type of plates are recommended for use with this kit?

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

    does the kit contain probenecid ?
    probenecid inactivates ion-transporters.
    customer usually adds it if component is not present in a Fluo-8 based kit, but if it is contained in the kit, he would not add it.


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

    Hello. I am writing about the Fluo-8 No Wash Calcium Assay Kit (ab112129). Mostly, I am wondering if it can be used with my Synergy 2 microplate reader from BioTek. The reader is set up with Ex/Em filters for 485/528, which seems OK, but it does not have ...

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    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 inc...

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    can kit be used with other species than human?


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

    Questions shown with answers below

    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, how...

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    We are wondering, what makes it no wash? How does one prevent that the cash will fluoresce outside the cells?

    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 ne...

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    Thanks for the prompt reply! I looked at other companies that offer Fluo-8 as well and the Kd depends on the chemistry of the molecule. Depending on the derivative it can range from 389 nM to as high as 10 uM, so the 230 nM sounds like it may not be the...

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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 389nm....

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    What is the Kd value of kit ab112129?

    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. Do...

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    Hi!

    I'm interested in the product: ab112115. My intention is to use it to measure a gradient of Ca+2 by fluorescent microscopy and study cell chimotaxis at the same time. I would like to know if this product can work in this conditions: physiolog...

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    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 Medi...

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    what is the KD value for binding to calcium?

    Thank you for contacting us. The kd is 320 nM.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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