Fluo-8 Calcium Flux Assay试剂盒- No Wash (ab112129)
Key features and details
- Assay type: Cell-based
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Adherent cells, Suspension cells
概述
-
产品名称
Fluo-8 Calcium Flux Assay试剂盒- No Wash
参阅全部 Calcium 试剂盒 -
检测方法
Fluorescent -
样品类型
Adherent cells, Suspension cells -
检测类型
Cell-based -
检测时间
1h 00m -
产品概述
Fluo-8 Calcium Flux Assay Kit (ab112129) is a no-wash, fluorescence-based assay for detecting intracellular calcium mobilization in cells from mammals and other species.
In the calcium flux assay 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.
This calcium flux assay provides an optimized assay method for monitoring 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.
Calcium flux assay protocol summary:
- add Fluo-8 dye solution to cells
- incubate for 30 min at 37ºC, and then for 30 min at room temp
- analyze fluoresence 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:
- No wash kit Fluo-8 Calcium Flux Assay Kit 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 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 No Wash Fluo-8 kit ab112129.
Fluo-8 is also available as a standalone biochemical ab142773.
-
说明
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).
-
平台
Microplate reader
性能
-
存放说明
Store at -20°C. Please refer to protocols. -
组件 96 tests 10X Pluronic® F127 Plus 1 x 1ml Fluo-8 1 vial HHBS 1 x 9ml -
研究领域
-
相关性
Calcium 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. -
别名
- Ca++
- Ca2+
相关产品
-
Positive Controls
-
Related Products
图片
-
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.
数据表及文件
-
Datasheet download
文献 (42)
ab112129 被引用在 42 文献中.
- Xing L et al. Critical role of mitogen-inducible gene 6 in restraining endothelial cell permeability to maintain vascular homeostasis. J Cell Commun Signal 17:151-165 (2023). PubMed: 36284029
- Alharthi J et al. A metabolic associated fatty liver disease risk variant in MBOAT7 regulates toll like receptor induced outcomes. Nat Commun 13:7430 (2022). PubMed: 36473860
- Noitem R et al. Natural statin derivatives as potential therapy to reduce intestinal fluid loss in cholera. PLoS Negl Trop Dis 16:e0010989 (2022). PubMed: 36490300
- Torz L et al. NPFF Decreases Activity of Human Arcuate NPY Neurons: A Study in Embryonic-Stem-Cell-Derived Model. Int J Mol Sci 23:N/A (2022). PubMed: 35328681
- Nakamichi R et al. The mechanosensitive ion channel PIEZO1 is expressed in tendons and regulates physical performance. Sci Transl Med 14:eabj5557 (2022). PubMed: 35648809