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Molecular Probes™ FluoVolt™ Membrane Potential Kit

Catalog No. F10488
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Description

FluoVolt™ Membrane Potential Kit

The FluoVolt™ Membrane Potential Kit represents the next generation in voltage sensitive fluorescent probe-based kits. The FluoVolt™ probe brings together the best characteristics of the fast and slow response membrane potential fluorescent probes. The FluoVolt™ probe responds to changes in membrane potential in sub-milliseconds and displays a high magnitude of response.

Features of the FluoVolt™ Membrane Potential Probe include:

  • Fast—responds to changes in membrane potential within sub-milliseconds

  • High sensitivity—response range is typically 25% per 100 mV

  • Emission/excitation works with standard FITC settings

  • Can be used in imaging or patch clamp applications

    Changes in membrane potential play a central role in many physiological processes, including nerve-impulse propagation, muscle contraction, and cell signaling. Potentiometric fluorescent probes are important tools for studying these processes and are generally characterized as slow- or fast-response probes.

    The Best Characteristics of Slow- and Fast-Response Probes

    Slow-response probes function by entering depolarized cells, binding to proteins or membranes, and exhibiting enhanced fluorescence. This membrane translocation event decreases the ability of these reporters to respond to changes in membrane potential and introduces a capacitive load that can affect cell health. However, these probes display a high magnitude of response; typically in the 1% per mV range.

    Fast-response probes are molecules that change their structure in response to the surrounding electric field and detect transient (millisecond) potential changes. However, when compared to the slow-response probes, the fast-response probes have a magnitude of potential-dependent fluorescence change that is often small (2–10% fluorescence change per 100 mV).

    The FluoVolt™ Membrane Potential Probe displays the best properties of the slow- and fast-response probes. The FluoVolt™ probe is a fast-response probe with a superior potential-dependent fluorescence response. The response is fast enough to detect transient (sub-millisecond) potential changes in excitable cells and generates a signal change in excess of 25% per 100 mV.

    PowerLoad™ and Background Suppressor Solutions Also Included

    For easy cell loading, the FluoVolt™ Membrane Potential Kit contains PowerLoad™ Concentrate. Due to the unique nature of the PowerLoad™ solution, it can be used in the presence of complete culture media, thus reducing the negative effects of replacing media or loading in serum-free media.

    Baseline autofluorescence caused by components within growth media can be greatly reduced by the addition of the included Neuro Background Supressor. This solution has been specifically formulated for use with neuronal cells and will not cause osmotic shock. Additionally, the Neuro Background Suppressor has been used successfully with many different cell types to efficiently suppress background fluorescence without sacrificing the specific cellular fluorescence generated in the assay.

    • Specifications

    Content And Storage Contains:
    • 50 μL of FluoVolt™ Dye
    • 500 μL of PowerLoad™ Concentrate
    • 5 mL of Neuro Background Suppressor
    • Store at 2°C to 8°C
    • Do not freeze
    Product Type Stain
    Product Line Molecular Probes
    Quantity 1 Kit
    Sub Cellular Localization Cell Membranes & Lipids

    Frequently Asked Questions (FAQs)

    I am seeing high background outside of my neuronal cells when using membrane potential indicators. What can I do to reduce background?

    If you use our FluoVolt Membrane Potential Kit (Cat. No. F10488), the kit provides a background suppressor to reduce this problem. For other indicators, consider the use of BackDrop Background Suppressor (Cat no. R37603, B10511, and B10512).

    Why do I lose all signal from my neuronal tracer when I do a methanol fixation on my cells?

    If the tracer you chose is a lipophilic dye and fix with methanol, the lipids are lost with the methanol. If you have to use methanol fixation then choose a tracer that will covalently bind to proteins in the neurons.
    I stained my cells with a lipophilic cyanine dye, like DiI, but the signal was lost when I tried to follow up with antibody labeling. Why?

    Since these dyes insert into lipid membranes, any disruption of the membranes leads to loss of the dye. This includes permeabilization with detergents like Triton X-100 or organic solvents like methanol. Permeabilization is necessary for intracellular antibody labeling, leading to loss of the dye. Instead, a reactive dye such as CFDA SE should be used to allow for covalent attachment to cellular components, thus providing for better retention upon fixation and permeabilization.
    I labeled my neurons with DiI and then fixed and permeabilized and now I have no signal. What did I do wrong?

    DiI is a lipophilic dye that resides mostly in lipids in the cell, when cells are permeabilized with detergent or fixed using alcohol this strips away the lipid and the dye. If permeabilization is required CM-DiI can be used because this binds covalently to proteins in the membrane; some signal is lost upon fixation/permeabilization, but enough signal should be retained to make detection possible.
    What is the difference between fast and slow-response membrane potential probes?

    Molecules that change their structure in response to the surrounding electric field can function as fast-response probes for the detection of transient (millisecond) potential changes. Slow-response dyes function by entering depolarized cells and binding to proteins or membranes. Increased depolarization results in additional dye influx and an increase in fluorescence, while hyperpolarization is indicated by a decrease in fluorescence. Fast-response probes are commonly used to image electrical activity from intact heart tissues or measure membrane potential changes in response to pharmacological stimuli. Slow-responding probes are often used to explore mitochondrial function and cell viability.
    What type of membrane potential indicators do you offer and how should I choose one for my experiment?

    A membrane potential indicator selection guide can be found here (https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-viability-and-regulation/ion-indicators/membrane-potential-indicators.html).
    Is there a way to label individual neurons without microinjecting?

    The solid and crystalline forms of DiI and other related dyes (Cat. Nos. D282, D3911, D7757, and D12731) are sometimes placed in contact with a specific neuron where it will travel down the cell by lateral diffusion via the membrane. Alternatively, our NeuroTrace Tissue Labeling Paste can be scooped onto a needle and placed onto particular neurons.

    Please see the information below for a comparison of our neuronal cell labeling methods:
    Product:Method of labeling: Labeling intensity: Features
    Neuron-specific antibodies: Primary antibodies directed to proteins expressed in neuronal cells: Proportional to the amount of protein expressed: Provides the only neuronal specific labeling method
    Lipophilic neuronal ytracers: Hydrophobic dyes are incorporated into lipids in the cell: This labeling method provides the most intense labeling becuase of the abundant amount of lipids: Allows tracing of neurons throughout the sample
    Membrane potential indicators: Dyes are loaded into live cells in aqueous buffers: Depends on either changes in structures due to the electrical field they are in, or dye influx due to depolarization: Changes in membrane potential play a central role in physiological processes, including nerve-impulse propagation, muscle contraction, and cell signaling

    For Research Use Only. Not for use in diagnostic procedures.