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AB112156

Rhod-4 Calcium Assay kit - Medium Removal No Wash

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(1 Publication)

Abcam Rhod-4 Calcium Assay kit - Medium Removal No Wash (ab112156) is a fluorescence-based assay for detecting intracellular calcium mobilization.
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Functional Studies - Rhod-4 Calcium Assay kit - Medium Removal No Wash (AB112156)
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Supplier Data

Functional Studies - Rhod-4 Calcium Assay kit - Medium Removal No Wash (AB112156)

Carbachol Dose Response measured in HEK293 cells using Rhod-4 Medium Removal No Wash Calcium Assay kit and Rhod-2.

HEK293 cells were incubated with 100 µL of the dye-loading solution using ab112156, or 100 µL of Rhod-2 solution (5 µM) for 1 hour at room temperature. Carbachol (25 µL/well) was added to achieve the final indicated concentrations. The EC50 of Rhod-4 is about 0.8 µM, and that for Rhod-2 about 0.7 µM.

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Results type

Quantitative

Assay time

1h

Assay Platform

Microplate reader

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Product details

Abcam Rhod-4 Calcium Assay kit - Medium Removal No Wash (ab112156) is a fluorescence-based assay for detecting intracellular calcium mobilization. Cells expressing a GPCR of interest that signals through calcium are pre-loaded with Rhod-4 which can cross the cell membrane. Once inside the cell, the lipophilic blocking groups of Rhod-4 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 Rhod-4. The characteristics of its long wavelength, high sensitivity, and >250 times fluorescence enhancement make Rhod-4 the brightest red calcium indicator available in the marker, and it is an ideal tool for the measurement of cellular calcium through HTS screening. Compared to Fluo-8, Rhod-4 is more photostable, making its fluorescence imaging more robust. ab112156 provides an optimized assay method for monitoring the 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. Visit our for tips and troubleshooting.

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.

If you would like to quantify calcium concentration in vitro using cell extracts, we recommend using Calcium Detection Kit (Colorimetric) (ab102505) Calcium Quantification Assay Kit (ab112115), as they provide stock standards for comparison.

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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C
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Product protocols

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Target data

See full target information Calcium
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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Cancer discovery 13:2270-2291 PubMed37553760

2023

GABA Regulates Electrical Activity and Tumor Initiation in Melanoma.

Applications

Unspecified application

Species

Unspecified reactive species

Mohita Tagore,Emiliano Hergenreder,Sarah C Perlee,Nelly M Cruz,Laura Menocal,Shruthy Suresh,Eric Chan,Maayan Baron,Stephanie Melendez,Asim Dave,Walid K Chatila,Jeremie Nsengimana,Richard P Koche,Travis J Hollmann,Trey Ideker,Lorenz Studer,Andrea Schietinger,Richard M White
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websiteProtocolBooklet
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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