FluoVolt™ Membrane Potential Kit
FluoVolt™ Membrane Potential Kit
Invitrogen™

FluoVolt™ Membrane Potential Kit

The FluoVolt™ Membrane Potential Kit represents the next generation in voltage sensitive fluorescent probe-based kits. The FluoVolt™ probe brings togetherLeia mais
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Número do catálogoQuantity
F104881 Kit
Número do catálogo F10488
Preço (BRL)
5.965,27
Each
Adicionar ao carrinho
Quantity:
1 Kit
Preço (BRL)
5.965,27
Each
Adicionar ao carrinho
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.

Learn more about ion indicators including calcium, potassium, pH, and membrane potential indicators ›

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.
For Research Use Only. Not for use in diagnostic procedures.
Especificações
Quantity1 Kit
Sub Cellular LocalizationCell Membranes & Lipids
Product LineMolecular Probes
Product TypeStain
Unit SizeEach
Conteúdo e armazenamento
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

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).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

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.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Citações e referências (12)

Citações e referências
Abstract
Screening fluorescent voltage indicators with spontaneously spiking HEK cells.
Authors:Park J, Werley CA, Venkatachalam V, Kralj JM, Dib-Hajj SD, Waxman SG, Cohen AE,
Journal:
PubMed ID:24391999
Development of improved fluorescent voltage indicators is a key challenge in neuroscience, but progress has been hampered by the low throughput of patch-clamp characterization. We introduce a line of non-fluorescent HEK cells that stably express NaV 1.3 and KIR 2.1 and generate spontaneous electrical action potentials. These cells enable rapid, ... More
Electrophysiology of hiPSC-Cardiomyocytes Co-Cultured with HEK Cells Expressing the Inward Rectifier Channel.
Authors:
Journal:Int J Mol Sci
PubMed ID:34205607
Patient-specific, re-engineered cardiomyocyte model confirms the circumstance-dependent arrhythmia risk associated with the African-specific common SCN5A polymorphism p.S1103Y: Implications for the increased sudden deaths observed in black individuals during the COVID-19 pandemic.
Authors:
Journal:Heart Rhythm
PubMed ID:34979239
hiPSC-CM Monolayer Maturation State Determines Drug Responsiveness in High Throughput Pro-Arrhythmia Screen.
Authors:
Journal:Sci Rep
PubMed ID:29061979
High-Throughput Drug Screening System Based on Human Induced Pluripotent Stem Cell-Derived Atrial Myocytes ∼ A Novel Platform to Detect Cardiac Toxicity for Atrial Arrhythmias.
Authors:
Journal:Front Pharmacol
PubMed ID:34413773