Abcam, ab189819, Membrane fluidity kit

Size: 100Test
Membrane Fluidity Assay Kit (ab189819) uses a fluorescent lipophilic pyrene probe that incorporates into the cell membrane. Upon spatial interaction and homodimer (or excimer) formation, the pyrene probe shifts its emission from Em ~400 nm to Em ~470 nm. Measuring the fluorescence ratio of Em ~470 nm to Em ~400 nm gives a quantitative measure of membrane fluidity. - Uses pyrenedecanoic acid (PDA) as probe - readout by flow cytometer and microscopy - Cited in over 35 publications
Key facts
Detection method:Fluorescent,
Assay Platform:Flow cytometer, Fluorescence microscope

Product details:
How the assay works
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. 400 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.
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.
Membrane fluidity kit protocol
Kit size: 100 tests in a 50 μL reaction volume.
1. Plate adherent cells or suspension cell
2. Prepare fluorescent labeling solution
3. Incubate cells with Fluorescent Lipid Reagent for 1 hour
4. Wash cells, resuspend cell in media
5. Read fluorescence at 400nm and 450-470nm
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.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C