Overview

  • Product name

    Cal-520, AM
  • General notes

    ab171868 provides a robust homogeneous fluorescence-based assay tool for detecting intracellular calcium mobilization. Cal-520 AM is a new fluorogenic calcium-sensitive dye with a significantly improved signal to noise ratio and intracellular retention compared to the existing green calcium indicators (such as Fluo-3 AM and Fluo-4 AM). Cells expressing a GPCR or calcium channel of interest that signals through calcium can be preloaded with Cal-520 AM which can cross cell membrane. Once inside the cell, the lipophilic blocking groups of Cal 520 AM are cleaved by esterases, resulting in a negatively charged fluorescent dye that stays inside cells. Its fluorescence is greatly enhanced upon binding to calcium. When cells stimulated with agonists, the receptor signals the release of intracellular calcium, which significantly increase the fluorescence of Cal-520. The characteristics of its long wavelength, high sensitivity, and >100 times fluorescence enhancement, make Cal-520 AM an ideal indicator for the measurement of cellular calcium. The high S/N ratio and better intracellular retention make the Cal-520 calcium assay a robust tool for evaluating GPCR and calcium channel targets as well as for screening their agonists and antagonists.

    Molecular Weight (MW) 1102.95 Da.

Properties

Images

  • CHO-M1 cells were seeded overnight at 40,000 cells per 100 μL per well in a 96-well black wall/clear bottom plate. 100 μl of 4 μM Cal 520 AM in HHBS were added into the wells, and the cells were incubated at 37 °C for 2 hour.

Protocols

References

This product has been referenced in:

  • Dave A  et al. Mitochondrial Electron Transport Chain Complex Dysfunction in MeCP2 Knock-Down Astrocytes: Protective Effects of Quercetin Hydrate. J Mol Neurosci 67:16-27 (2019). Read more (PubMed: 30519865) »
  • Bai L  et al. A Conserved Circadian Function for the Neurofibromatosis 1 Gene. Cell Rep 22:3416-3426 (2018). Read more (PubMed: 29590612) »
See all 3 Publications for this product

Customer reviews and Q&As

Very good signal

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Cal-520 used for calcium imaging on primary astrocyte culture.
Used following the protocol provided and imaged under a Leica Sp6.

Figure included is an example of image when astrocytes are stimulated with ATP (50uM).

Ms. Maria Velasco

Verified customer

Submitted Jun 05 2018

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