Key features and details
- Detection method: Fluorescent
- Platform: Flow cytometer, Fluorescence microscope
Product nameMembrane fluidity kit
Membrane fluidity or "membrane viscosity" for short range lateral diffusion has best been measured using lipid analog probes that, when interacting, exhibit changes in their spectral properties. One of the best systems for use in such studies are the lipophilic pyrene probes that undergo excimer formation upon spatial interaction. When excimers form, the emission spectrum of the pyrene probe shifts dramatically to the red (longer wavelength). By measuring the ratio of monomer (EM max. 372 nm) to excimer (EM 470 nm) fluorescence, a quantitative monitoring of the membrane fluidity can be attained. These measurements can provide kinetic information, as well as in vivo monitoring of cellular function by both flow cytometry and microscopic analysis.
Exitation wavelength (nm): 350
Emission wavelength (nm): 450
The dynamic properties of the cell membrane and cytoplasmic microtubules and microfilaments, as well as the dynamic movement of lipids in micelles and vesicles is of importance in such diverse areas as activation of polymorphonuclear leukocytes and chemotaxis, activation of membrane enzyme systems and the specific assembly or mobilization of microtubules and microfilaments, enhancement of the affinity of chemoattractant receptors, as well as being associated with a variety of pathological syndromes related to membrane fluidity.
It has been recognized that the rotational mobility of fluorescent or magnetic resonant probes is different from that observed in lateral diffusion.
Kit size: 100 tests in a 50 µL reaction volume.
PlatformFlow cytometer, Fluorescence microscope
Components 100 tests Fluorescent Lipid Reagent 1 x 2ml Perfusion Buffer 1 x 25ml Pluronic F-127 1 x 50mg Reference Standard 1 x 2ml
Isopropanol treatment increases membrane fluidity in primary leukocytes
Primary leukocytes were treated with .5% isopropanol for 1 hour, followed by 1 hour incubation with 5 µM PDA and 0.08% pluronic F-127. PDA fluorescence was measured by exciting at 350 nM and taking emission values at 400nM (monomer) and 460nM (excimer). Relative membrane fluidity is a ratio of excimer to monomer fluorescence.
Ethanol treatment increases membrane fluidity in HeLa cells
HeLa cells were treated with 100mM or 200 mM Ethanol for 48 hours, followed by 1 hour incubation with 2 µM PDA and 0.08% pluronic F-127. PDA fluorescence was monitored by exciting at 350 nM and taking emission values at 400nM (monomer) and 470nM (excimer). Relative membrane fluidity is a ratio of excimer to monomer fluorescence. Aliphatic alcohols such as EtOH are known to increase membrane fluidity.
Gaucher Disease leukocytes exhibit decreased membrane fluidity relative to healthy controls
Immortalized leukocytes were incubated for 1 hour incubation with 2 µM PDA and 0.08% pluronic F-127. PDA fluorescence was measured by exciting at 350nM and taking emission values at 400nM (monomer) and 470nM (excimer). Relative membrane fluidity is a ratio of excimer to monomer fluorescence. Previous studies have demonstrated that cells isolated from Gaucher Disease patients exhibit decreased membrane fluidity.
ab189819 has been referenced in 26 publications.
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- Piccarducci R et al. Impact of ApoE Polymorphism and Physical Activity on Plasma Antioxidant Capability and Erythrocyte Membranes. Antioxidants (Basel) 8:N/A (2019). PubMed: 31717561