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Cell Viability Assay Kit (Fluorometric - Green) (ab112122)

Submit a review Q&A (3)References (1)
Functional Studies - Cell Viability Assay Kit (Fluorometric - Green) (ab112122)

    Key features and details

    • Assay type: Quantitative
    • Detection method: Fluorescent
    • Platform: Microplate reader, Fluor. microscope, Flow cyt.
    • Sample type: Adherent cells, Suspension cells

    Overview

    • Product name

      Cell Viability Assay Kit (Fluorometric - Green)
      See all Cell Viability kits

    • Detection method

      Fluorescent

    • Sample type

      Adherent cells, Suspension cells

    • Assay type

      Quantitative

    • Species reactivity

      Reacts with: Mammals, Other species

    • Product overview

      There are a variety of parameters that can be used to monitor cell viability. The green fluorescent dye used in the kit is a hydrophobic compound. It easily permeates intact live cells and gets enhanced fluorescence upon entering into live cells. The hydrolysis of the non-fluorescent substrate by intracellular esterases generates a strongly green fluorescent hydrophilic product that is well-retained in the cell cytoplasm. The green fluorophore generated by the non-fluorescent substrate used in the kit has the spectral properties of fluorescein at Ex/Em = ~490 nm/520 nm. When well excited with the Argon Laser at 488 nm, the fluorophore emits intense green fluorescence at ~520 nm.


      ab112122 provides all the essential components with an optimized cell-labeling protocol for fluorescence microplate assays. It can also be used with a fluorescence microscope equipped with a FITC filter set.


      ab112122 provides an effective tool of labeling cells for fluorescence microplate and microscopic investigations of cellular functions. It is useful for a variety of studies, including cell adhesion, chemotaxis, multidrug resistance, cell viability, apoptosis and cytotoxicity. ab112122 is suitable for proliferating and non-proliferating cells.

      Visit our FAQs page for tips and troubleshooting.

    • Notes

      Review the cell health assay guide to learn about more kits to perform a cell viability assay, cytotoxicity assay and cell proliferation assay. 

    • Platform

      Microplate reader, Fluor. microscope, Flow cyt.

    Properties

    Images

    • Functional Studies - Cell Viability Assay Kit (Fluorometric - Green) (ab112122)
      Functional Studies - Cell Viability Assay Kit (Fluorometric - Green) (ab112122)
      CHO-K1 cell number response was measured with ab112122. CHO-K1 cells at 0 to 5,000 cells/well/100 µL were seeded overnight in a black wall/clear bottom 96-well plate. The cells were incubated with 100 µL/well of Green dye-loading solution for 1 hour at 37 °C. The fluorescence intensity was measured at Ex/Em = 490/ 525 nm. The fluorescence intensity was linear (R2 = 1) to the cell number as indicated. The detection limit was 30 cells/well (n=6).

    References (1)

    Publishing research using ab112122? Please let us know so that we can cite the reference in this datasheet.

    ab112122 has been referenced in 1 publication.

    • Kanno S  et al. Establishment of a developmental toxicity assay based on human iPSC reporter to detect FGF signal disruption. iScience 25:103770 (2022). PubMed: 35146387

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    Question

    What are the names of the fluorophores in the kits ab112121 and ab112122 ?

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    Answer



    Les filtres à utiliser sont FITC (vert)et Cy3ou Texas red (rouge).

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    Question

    Hi,

    I am using Greiner Thincert Culture Inserts for 6-well plates and A549 cells in my exposure experiments. The goal is to expose the cells at an air/liquid interphase for a certain period and to measure cell viability, oxidative stress markers and cell cycle stage after the exposure.

    Until now I have tried to detach the cells from the inserts and measure the MTT assay and do flow cytometry analysis. However I have been having problems detaching the cells from the inserts, trypsin/EDTA is ineffective and scraping destroys the cells, so I have not been able to get sensible data from our exposure experiments.

    Can you recommend any kits/procedures which could be applied directly in the inserts without prior detachments of the cells?

    The minimum parameters I would need to evaluate after the exposure are cell viability and apoptosis/cell cycle stage, but I would like to include a marker for oxidative stress (e.g. glutathione) as well. I should be able to measure all the mentioned parameters simultaneously in one insert without too much quenching and fluorescence interference/overlay.

    We have a Victor III microplate reader and an inverted fluorescence microscope for which we will acquire quantification software shortly.

    Thank you for your help.

    With best regards,

    Read More
    Answer

    Thank you for your enquiry and interest in our products.

    MTT assay can be carried out on 96 well plates using either adherent or floating cells. You do not necessary need to remove (harvest) the cells by trypsinization. You can culture the cells in the wells of the 96 well plates, then apply the MTT solution (20ul) to each well. During incubation (37C, 5% CO2) for 1-5 hours MTT will be metabolized. The formazan (MTT metabolic product) can then be dissolved in DMSO and the optical density can be read at 560 nm using a plate reader - without harvesting the cells. There may be other cell viability/cytotoxicyprotovols but this is the most widely used.

    Cell viability kits:

    We have different cell viability assay kits in our catalogue i.e. ab112120, ab112121, ab112122, ab112123.

    https://www.abcam.com/Cell-Viability-Assay-Kit-Fluorometric-Blue-ab112120.html

    https://www.abcam.com/Cell-Viability-Assay-Kit-Fluorometric-Dual-Green-Red-ab112121.html

    https://www.abcam.com/Cell-Viability-Assay-Kit-Fluorometric-Green-ab112122.html

    https://www.abcam.com/Cell-Viability-Assay-Kit-Fluorometric-Near-InfraRed-ab112123.html

    GSH kits:

    GSH can be detected/quantified either by fluorimetric or chromogenic (DTNB - Ellman's reagent) assays and currently we have also some assay kits for detecting this tripeptide, you may wish to take a look at the datasheets for further information.

    - ab65322: Glutathione Fluorometric Detection Kit

    https://www.abcam.com/Glutathione-Fluorometric-Detection-Kit-ab65322.html

    - ab112132:Intracellular GSH Assay Kit

    https://www.abcam.com/Intracellular-GSH-Assay-Kit-ab112132.html

    Normally, intracellular GSH levels can be expressed per cell numbers or per protein concentration.

    I hope this helps and if I can assist further, please do not hesitate to contact me.

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    Question

    I have recently purchased the abcam Fluorescent Green Cell Viability Assay (#ab112122).  I have have trouble conducting the assay.  I am using L929 fibroblasts (5k per well in 96-well plates) in a standard ISO 10993 cytotoxicity assay.  I have prepared the loading solution according to the instructions and added 100uL of the loading solution to each well containing cells and treatment extracts.  I read the plate at 490/525/nm.  The controls are not at the expected levels.  Furthermore, a set of blank wells containing only media (EMEM + serum) showed very high background (below).   Can you provide some insight on the difficulty I am having with this assay?  The level of variation in the blank concerns me.  

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    Answer

    Thank you for contacting us regarding this issue. I have spoken with the lab about you results using ab112122. It seems from your results that most likely you have used the top read mode. For this assay, it must use bottom read mode for measuring, and also use cutoff filter at 515nm if using Flexstation from Molecular Devices. We hope this helps you. Please do not hesitate to contact us if you have other questions or if there are more ways that we may help you reach your research goals.

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