Calcium Flux Assay Kit (Fluo-8, No Wash) ab112129 is a no-wash, fluorescence-based assay for detecting intracellular calcium mobilization in cells from mammals and other species.
Fluo-8 Calcium Flux Assay Kit - No Wash designed for success:
- Ideal indicator for measurement of cellular calcium because of its long wavelength, high sensitivity, and 100-250 times fluorescence increases (when it forms complexes with calcium)
- Easy to use and fast protocol with no wash step required
- Can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation
- Cited in > 50 publications
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- Nature 625:557-5652024Na1.7 as a chondrocyte regulator and therapeutic target for osteoarthritis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWenyu Fu,Dmytro Vasylyev,Yufei Bi,Mingshuang Zhang,Guodong Sun,Asya Khleborodova,Guiwu Huang,Libo Zhao,Renpeng Zhou,Yonggang Li,Shujun Liu,Xianyi Cai,Wenjun He,Min Cui,Xiangli Zhao,Aubryanna Hettinghouse,Julia Good,Ellen Kim,Eric Strauss,Philipp Leucht,Ran Schwarzkopf,Edward X Guo,Jonathan Samuels,Wenhuo Hu,Mukundan Attur,Stephen G Waxman,Chuan-Ju LiuPubMed 38172636
- International journal of molecular sciences 24:2023MELAS-Derived Neurons Functionally Improve by Mitochondrial Transfer from Highly Purified Mesenchymal Stem Cells (REC).Applications:Unspecified applicationReactive species:Unspecified reactive speciesLu Liu,Jiahao Yang,Yoshinori Otani,Takahiro Shiga,Akihiro Yamaguchi,Yasuaki Oda,Miho Hattori,Tsukimi Goto,Shuichi Ishibashi,Yuki Kawashima-Sonoyama,Takaya Ishihara,Yumi Matsuzaki,Wado Akamatsu,Masashi Fujitani,Takeshi TaketaniPubMed 38139018
- Nature cell biology 25:1774-17862023NAD regulates nucleotide metabolism and genomic DNA replication.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSebastian Howen Nesgaard Munk,Joanna Maria Merchut-Maya,Alba Adelantado Rubio,Arnaldur Hall,George Pappas,Giacomo Milletti,MyungHee Lee,Lea Giørtz Johnsen,Per Guldberg,Jiri Bartek,Apolinar Maya-MendozaPubMed 37957325
- The journal of headache and pain 24:1412023The potent analgesia of intrathecal 2R, 6R-HNK via TRPA1 inhibition in LF-PENS-induced chronic primary pain model.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAn-Ran Liu,Zhen-Jia Lin,Ming Wei,Yuan Tang,Hui Zhang,Xiang-Ge Peng,Ying Li,Yu-Fan Zheng,Zhi Tan,Li-Jun Zhou,Xia FengPubMed 37858040
- Frontiers in immunology 14:11066932023IP3R-dependent mitochondrial dysfunction mediates C5b-9-induced ferroptosis in trichloroethylene-caused immune kidney injury.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZhibing Liu,Jinru Ma,Xulei Zuo,Xuesong Zhang,Haibo Xie,Feng Wang,Changhao Wu,Jiaxiang Zhang,Qixing ZhuPubMed 37383224
- PLoS pathogens 19:e10112952023Porcine reproductive and respiratory syndrome virus infection triggers autophagy via ER stress-induced calcium signaling to facilitate virus replication.Applications:Unspecified applicationReactive species:Unspecified reactive speciesFeifei Diao,Chenlong Jiang,Yangyang Sun,Yanni Gao,Juan Bai,Hans Nauwynck,Xianwei Wang,Yuanqi Yang,Ping Jiang,Xing LiuPubMed 36972295
- STAR protocols 4:1021152023Protocol for high-throughput single-cell patterning using a reusable ultrathin metal microstencil.Applications:Unspecified applicationReactive species:Unspecified reactive speciesQingqing Tian,Kunming Xing,Yongshu Liu,Qian Wang,Haonan Sun,Ying-Nan Sun,Shusheng ZhangPubMed 36853712
- PLoS neglected tropical diseases 16:e00109892022Natural statin derivatives as potential therapy to reduce intestinal fluid loss in cholera.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRattikarn Noitem,Pawin Pongkorpsakol,Chartchai Changsen,Yaowapa Sukpondma,Chittreeya Tansakul,Vatcharin Rukachaisirikul,Chatchai MuanprasatPubMed 36490300
- Nature communications 13:74302022A metabolic associated fatty liver disease risk variant in MBOAT7 regulates toll like receptor induced outcomes.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJawaher Alharthi,Ali Bayoumi,Khaled Thabet,Ziyan Pan,Brian S Gloss,Olivier Latchoumanin,Mischa Lundberg,Natalie A Twine,Duncan McLeod,Shafi Alenizi,Leon A Adams,Martin Weltman,Thomas Berg,Christopher Liddle,Jacob George,Mohammed EslamPubMed 36473860
- Journal of cell communication and signaling 17:151-1652022Critical role of mitogen-inducible gene 6 in restraining endothelial cell permeability to maintain vascular homeostasis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLiying Xing,Guanqun Huang,Rongyuan Chen,Lijuan Huang,Juanxi Liu,Xiangrong Ren,Shasha Wang,Haiqing Kuang,Anil Kumar,Jong Kyong Kim,Qin Jiang,Xuri Li,Chunsik LeePubMed 36284029
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Cell-based
- Assay time
- 1h
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Target data
Alternative names
Ca++, Ca2+
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Calcium Flux Assay Kit (Fluo-8, No Wash) ab112129 is a no-wash, fluorescence-based assay for detecting intracellular calcium mobilization in cells from mammals and other species.
Fluo-8 Calcium Flux Assay Kit - No Wash designed for success:
- Ideal indicator for measurement of cellular calcium because of its long wavelength, high sensitivity, and 100-250 times fluorescence increases (when it forms complexes with calcium)
- Easy to use and fast protocol with no wash step required
- Can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation
- Cited in > 50 publications
Key facts
- Detection method
- Fluorescent
- Sample types
- Suspension cells, Adherent cells
- Assay type
- Cell-based
- Assay time
- 1h
- Assay Platform
- Microplate reader
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
How the assay works
In the Fluo-8 Calcium Flux Assay Kit protocol, 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.
Fluo-8 Calcium Flux Assay Kit protocol Summary
1. Prepare cells in growth medium with 1-5% FBS
2. Add Fluo-8 NW dye-loading solution (100 µL/well for 96-well plate or 25 µL/well for 384-well plate)
3. Incubate at room temperature for 1 hour
4. Monitor fluorescence intensity at Ex/Em = 490/525 nm
Fluo-8 products available
Fluo-8 Calcium Flux Assay Kits are available in No Wash and Medium Removal variants:
- Calcium Flux Assay Kit (Fluo-8, No Wash) ab112129 is generally preferred as the workflow is simpler. It is also recommended for cell lines that are only weakly adherent and risk losing adherence when washing.
- Medium removal kit Fluo-8 Calcium Flux Assay Kit Fluo-8 Calcium Assay Kit - Medium Removal ab112128 is recommended for the testing of compounds, receptors and calcium channels that are sensitive to the presence of cell culture media or serum. Alternatively, for cells that can tolerate it, consider using a low serum % in culture media and use Calcium Flux Assay Kit (Fluo-8, No Wash) ab112129.
Fluo-8 is also available as a standalone biochemical Fluo-8 AM, green fluorescent calcium binding dye ab142773.
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) (Calcium Assay Kit (Colorimetric) ab102505) or Calcium Quantification Assay Kit (Calcium Assay kit (Fluorometric) ab112115).
How other researchers are using
Fluo-8 Calcium Flux Assay Kit - No Wash has been used in a variety of sample type including:
- Human OA chondrocytes and C28I2 cells 1
-Neuronal cells 2
- Human U2OS, HeLa, BJ, BJ-5ta (BJ-T), T98G, U87 MG and MRC5 cell lines and mouse embryonic fibroblasts (MEF) 3
References:
1- Fu W et al. 2024; 2- Lui L et al. 2023; 3- Munk S et al. 2023
Related and recommended products
See other alternative kits to quantify calcium concentration in vitro using cell extracts:
- Calcium Detection Kit (Colorimetric) Calcium Assay Kit (Colorimetric) ab102505
- Calcium Quantification Assay Kit Calcium Assay kit (Fluorometric) ab112115.
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Functional Studies - Calcium Flux Assay Kit (Fluo-8, No Wash) (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.
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