All products have special prices for bulk purchase, please contact us for more details if required.

Cat. No.: MCAKR2A-200 (for 20-200T)

Cat. No.: MCAKR2A-1k (for 100-1000T)

Description

Mitochondrial Calcium Assay Kit with Rhod-2 AM, also known as the Mitochondrial Calcium Influx Assay Kit with Rhod-2 AM, is a kit that uses Rhod-2 AM as a fluorescent probe to quickly and sensitively detect changes in mitochondrial calcium ions (Ca2+). The kit is compatible with various fluorescence detection systems such as fluorescence microscopes, laser confocal microscopes, fluorescent plate readers, and flow cytometers.

Calcium is typically the most abundant mineral in mammals, playing a crucial role in regulating numerous cellular activities and serving as one of the most important intracellular regulatory factors. Mitochondrial calcium ions regulate various mitochondrial functions, including oxidative metabolism and ATP synthesis, and are involved in cell death regulation. A moderate increase in mitochondrial calcium ions activates enzymes in the citric acid cycle, promoting ATP synthesis. However, an overload of mitochondrial calcium ions can lead to mitochondrial dysfunction and cell death.

Rhod-2 AM (also known as RHOD 2-AM, RHOD-2 AM, Mitochondrial Calcium Ion Fluorescent Probe), with CAS number 145037-81-6, has a molecular weight of 1124.0 and the molecular formula C52H59N4O19·Br. Rhod-2 AM is an acetoxymethyl ester derivative of Rhod-2 and is cell membrane permeable. Once inside the cell, it is hydrolyzed by intracellular esterases to form Rhod-2, which is then retained within the cell. Rhod-2's structure, similar to the mitochondrial-specific probe Rhodamine, allows it to enter mitochondria and bind to mitochondrial calcium ions. Upon binding with calcium ions, Rhod-2 exhibits strong fluorescence under 552nm excitation light, making it a common probe for monitoring mitochondrial calcium ion concentration changes.

Rhod-2 is somewhat similar to Fluo-3 and Fluo-4 in that its excitation and emission spectra do not shift significantly with changes in calcium ion concentration. However, unlike Fluo-3 and Fluo-4, Rhod-2 can specifically detect changes in mitochondrial calcium ions. Additionally, Rhod-2 has a longer wavelength, making it suitable for calcium ion detection experiments in cells and tissues with high levels of autofluorescence. It is also suitable for detecting calcium ions produced by visible light-activated calcium chelators and photoreceptors. The maximum excitation wavelength of Rhod-2 is 552nm, and the maximum emission wavelength is 581nm.

Advantage

Convenient Usage with Comprehensive Reagents: This kit includes several supporting reagents for user convenience. It provides an Assay Buffer for diluting Rhod-2 AM, which greatly ensures cell viability and maintains near-normal growth conditions. The Solubility Enhancer improves the staining effect by enhancing the solubility of Rhod-2 AM. The Staining Enhancer effectively maintains intracellular fluorescence and reduces background fluorescence during detection. Additionally, the kit includes CaUp as a positive control for inducing an increase in mitochondrial calcium ion concentration in cells.

Components

• Rhod-2 AM (500X)
• Solubility Enhancer (500X)
• Assay Buffer
• Staining Enhancer (100X)
• CaUp (500X)    

Storage

• Store at -20ºC for up to one year, except for Rhod-2 AM (500X).
• Store Rhod-2 AM (500X) at -80ºC.
• Both Rhod-2 AM (500X) and CaUp (500X) should be protected from light.

Precautions

• For the first use, aliquot appropriately to avoid repeated freeze-thaw cycles.
• Avoid using BSA and Phenol Red as they interfere with the detection of the fluorescent probe.
• This kit is only for detecting live cells or tissues, not for fixed or frozen samples. However, cells or tissues can be fixed after staining.
• For fluorescence plate reader detection, use plates suitable for fluorescence detection.
• Ensure Rhod-2 AM (500X) and Solubility Enhancer (500X) are fully dissolved before use.
• Fluorescent dyes are subject to quenching; avoid light exposure to minimize fluorescence quenching.
• This product is for scientific research by professionals only. It is not for clinical diagnosis or treatment, nor for use in food or medicine. Do not store it in common household environments.
• For your safety and health, wear a lab coat and disposable gloves when handling.