MarkerGene™ Membrane Fluidity Kit

Product ID: M0271



Unit SizePriceQuantity 
1kit
$145.42
  • Buy 5 for $116.34 each and save 20%

Availability: In stock


Description

This kit allows quantitative monitoring of membrane fluidity in cell membranes, micelles, and vesicles through use of a lipophilic pyrene probe.

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 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 excimer (EM 470 nm) to monomer (EM max. 372 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.


Technical Data
SKU M0271
Unit Size 1kit
Detection Method Fluorescence

References and Citations

Citations:

  • Saha AK, Dallo SF, Detmar AL, Osmulski P, Gaczynska M, Huang TH, Ramasubramanian AK. (2016) "Cellular cholesterol regulates monocyte deformation." J Biomech. pii: S0021-9290(16)31326-4.
  • Evani SJ, Ramasubramanian AK. Biophysical regulation of Chlamydia pneumoniae-infected monocyte recruitment to atherosclerotic foci. Sci Rep. 2016 Jan 20;6:19058.
  • Gao S, Guan Q, Chafeeva I, Brooks DE, Nguan CY, Kizhakkedathu JN, Du C.(2015) "Hyperbranched polyglycerol as a colloid in cold organ preservation solutions." PLoS One. 10(2):e0116595.
  • Guarnieri S, Morabito C, Belia S, Barberi L, Musarò A, Fanò-Illic G, Mariggiò MA. (2014) "New insights into the relationship between mIGF-1-induced hypertrophy and Ca2+ handling in differentiated satellite cells." PLoS One. 9(9):e107753.
  • Mendez TL, De Chatterjee A, Duarte TT, Gazos-Lopes F, Robles-Martinez L, Roy D, Sun J, Maldonado RA, Roychowdhury S, Almeida IC, Das S. (2013) "Glucosylceramide transferase activity is critical for encystation and viable cyst production by an intestinal protozoan, Giardia lamblia." J. Biol. Chem. 288(23):16747-16760.
  • Koeberle A, Shindou H, Harayama T, Yuki K, Shimizu T. (2012) "Polyunsaturated fatty acids are incorporated into maturating male mouse germ cells by lysophosphatidic acid acyltransferase 3." FASEB J. 26(1):169-180.
  • Tavolari S, Munarini A, Storci G, Laufer S, Chieco P, Guarnieri T. (2012) "The decrease of cell membrane fluidity by the non-steroidal anti-inflammatory drug Licofelone inhibits epidermal growth factor receptor signalling and triggers apoptosis in HCA-7 colon cancer cells." Cancer Lett. 321(2):187-194.
  • Guan Q, Li S, Yip G, Gleave ME, Nguan CY, Du C. (2012) "Decrease in donor heart injury by recombinant clusterin protein in cold preservation with University of Wisconsin solution." Surgery. 151(3):364-371
  • Li S, Guan Q, Chen Z, Gleave ME, Nguan CY, Du C.(2011) "Reduction of cold ischemia-reperfusion injury by graft-expressing clusterin in heart transplantation." J. Heart Lung Transplant. 30(7):819-826.
  • Azenabor AA, Job G, Adedokun OO. (2005) "Chlamydia pneumoniae infected macrophages exhibit enhanced plasma membrane fluidity and show increased adherence to endothelial cells." Mol. Cell. Biochem. 269(1-2):69-84
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References:

  • Dix JA, Verkman AS. (1990) "Pyrene eximer mapping in cultured fibroblasts by ratio imaging and time-resolved microscopy." Biochemistry 29(7): 1949-1953.
  • Masuda M, Kuriki H, Komiyama Y, Nishikado H, Egawa H, Murata K. (1987) "Measurement of membrane fluidity of polymorphonuclear leukocytes by flow cytometry." J. Immunol. Meth. 96(2): 225-231.
  • Neufeld ND, Corbo LM. (1984) "Membrane fluid properties of cord blood mononuclear leucocytes: association with increased insulin receptors." Pediatric Res. 18(8): 773-778.
  • Tomonaga A, Hirota M, Snyderman R. (1983) "Effect of membrane fluidizers on the number and affinity of chemotactic factor receptors on human polymorphonuclear leukocytes." Microbiol. Immunol. 27(11): 961-972.
  • Yuli I, Tomonaga A, Synderman R. (1982) "Chemoattractant receptor functions in human polymorphonuclear leukocytes are divergently altered by membrane fluidizers."Proc. Natl. Acad. Sci. USA 79(19): 5906-5910.
  • Haak RA, Ingraham LM, Baehner RL, Boxer LA. (1979) "Membrane fluidity in human and mouse Chediak-Higashi leukocytes." J. Clin Invest. 64(1): 138-144.
  • Oliver JM. (1978) "Cell biology of leukocyte abnormalities-membrane and cytoskeletal function in normal and defective cells. A review." Amer. J. Pathology 93(1): 221-270.
  • Galla HJ, Luisetti J. (1980) "Lateral and transversal diffusion and phase transitions in erythrocyte membranes. An excimer fluorescence study." Biochim Biophys Acta Biomembranes 596(1): 108-17.
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Technical Support

Question about this product? Ask a Scientist!

We pride ourselves on the high quality of our products and want you to get the best possible results from your assays. If you have any questions about this product or need help optimizing your protocol check out the product FAQs below or ask your own question and one of our expert scientists will get back to you asap:


⚛ Question: Can this kit be used for staining live, frozen or fixed tissue samples?
This method is usually appropriate for cell monolayers or cells grown in culture. It might be best to digest the tissues and measure the fluidity of the individual cells. But you could try to stain the live tissues directly (not fixed). We haven't tried to measure the fluidity in live tissue samples but you might be able to get it to work by bathing the tissues using the same protocol as described for cell monolayers. Application of known fluidity enhancing reagents (i.e. ethanol) or changes in temperature can be used to determine if the method(s) are working.
⚛ Question: Can I use this kit without a plate reader?
It is not necessary to have a polarized fluorescence spectrometer (for fluorescence polarization) in order to measure the eximer levels in cells. We were able to monitor eximer level increases with reduced temperature, in cells grown in culture using a fluorescence microscope. Reference for use with a microscope: Dix JA, Verkman AS. (1990) “Pyrene excimer mapping in cultured fibroblasts by ratio imaging and time-resolved microscopy.” Biochemistry 29: 1949-1953. Reference for use with Flow cytometry: Masuda M, Kuriki H, Komiyama Y, Nishikado H, Egawa H, Murata K. (1987) “Measurement of membrane fluidity of polymorphonuclear leukocytes by flow cytometry.” J Immunol Meth 96: 225 231.
⚛ Question: Is there a max/min cell count I should use?
I suggest 5000-10000 cells per well, depending on size. Usually I shoot for 60-80% confluency for my experiments. If you have to let the cells adhere overnight, you will need to adjust for the cells growing during that time period. Confluent cells often change their biology compared to actively growing cells.
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