All AbMole products are for research use only, cannot be used for human consumption.
Laurdan is a membrane-permeable fluorescent probe that is spectrally sensitive to the phospholipid phase of cell membranes to which it binds, and can be used for the identification of phospholipid phases, the absorption wavelength of Laurdan is 340-380nm and the emission wavelength is 440-490nm. Laurdan is phase-sensitive. Its fluorescence emission in the liquid crystal phase will show a large red shift, while it will show a small red shift in the gel phase, which can detect the phase state and fluidity of the membrane. Laurdan can measure the generalized polarization (GP) of the cell membrane. High GP is usually associated with low fluidity, low polarity or high cholesterol content of the membrane, while low GP is the opposite.
The following experimental protocol only provides a reference for a Laurdan generalized polarization (GP) measurement experiment of cell membrane. Please modify the working concentration and staining time according to experimental needs or refer to the literature. (from the literature, for reference only)
1. Solution preparation
(1) Storage solution: Dissolve Laurdan in DMSO or chloroform to a final concentration of 5 mM.
Note: After unused storage solution is aliquoted, store it at -20℃ in the dark to avoid repeated freezing and thawing.
(2) Working solution: Dilute the storage solution with experimental buffer (e.g. cell culture medium) to the required working concentration, usually in the range of 1-10 µM.
Note: Please adjust the optimal working concentration according to the actual situation or refer to the literature to set the gradient concentration by yourself. The working solution must be prepared and used immediately.
2. Steps for measuring the generalized polarization (GP) of Bacillus subtilis cell membrane by Laurdan
(1) Cell treatment: Cultivate cells to the middle of the logarithmic growth phase, wash them four times in PBS containing 2% glucose and 1% DMF, resuspend them in the same buffer, and adjust the OD600 to 0.3.
(2) Liposome preparation: Bacillus subtilis polar lipid extract was prepared by detergent dialysis, and the prepared liposome solution was squeezed through a 0.4μm filter membrane 20 times. The 10mg/ml liposome stock solution was diluted to a concentration of 1 mg/ml with Tris buffer (5mM, pH 7.4).
(3) Liposome labeling: Add Laurdan solution to the cell suspension to a final concentration of 10 μM and incubate in the dark for 30 minutes.
(4) Fluorescence measurement: Laurdan fluorescence measurement was performed using a BioTek Synergy MX plate reader. The excitation wavelength was 350 nm, the emission wavelengths were 460 nm and 500 nm, and readings were taken every 2 minutes.
(5) Laurdan fluorescence polarization calculation: Laurdan generalized polarization (GP) value was calculated using the following formula: (I460-I500)/(I460+I500).
References: https://pubmed.ncbi.nlm.nih.gov/29451901/
| Molecular Weight | 353.54 |
| Formula | C24H35NO |
| CAS Number | 74515-25-6 |
| Form | Solid |
| Solubility (25°C) | DMSO 5 mg/mL Chloroform 10 mg/mL |
| Storage | Powder 4°C, protect from light |
| Related Fluorescent Dye Products |
|---|
| Copper probe CF4
Copper probe CF4 (Copper fluor CF4) is a Cu+-specific fluorescent probe based on a rhodol dye scaffold. Copper probe CF4 (Copper fluor CF4) has high copper selectivity with a Kd value of 2.9×10-13 M, particularly over zinc and iron, as well as abundant cellular alkali and alkaline earth metals. Copper probe CF4 (Copper fluor CF4) is stable in a physiologically relevant pH regime between 6 and 8 (wavelengths of 415 nm for excitation and 660 nm for emission). Copper probe CF4 (Copper fluor CF4) can be used to study colon cancer. |
| Phalloidin-Fluor 647 Conjugate
Phalloidin-Fluor 647 Conjugate selectively binds to F-actins. Used at nanomolar concentrations, phalloidin derivatives are convenient probes for labeling, identifying and quantitating F-actins in formaldehyde-fixed and permeabilized tissue sections, cell cultures or cell-free experiments. |
| HBC530
HBC530 is a GFP fluorophore-like synthetic dye, with a structurally rigid electron acceptor and a strong electron donor. HBC is nonfluorescent in solution, and when combined with Pepper (RNA aptamer), HBC forms a tight complex and activates and emits bright fluorescence (Kd of ~3.5 nM). HBC emission peaks vary in different complexes and covers the spectrum from cyan to red. HBC can be used in the live cell imaging of RNA (Em/Ex = 530/485 nm). |
| 2',7'-dichlorodihydrofluorescein
2,7-Dichlorodihydrofluorescein (DCFH2) is a non-fluorescent reactive oxygen species (ROS) probe with an excitation wavelength of 485-500 nm and an emission wavelength of 515-530 nm. 2,7-Dichlorodihydrofluorescein is first hydrolyzed by intracellular esterases and then oxidized by ROS to generate non-biomembrane-permeable, highly fluorescent 2,7-dichlorofluorescein (DCF). 2,7-Dichlorodihydrofluorescein can undergo oxidation reactions with a variety of ROS (such as ·OH, H2O2, ONOO-, etc.) and is used to quantitatively detect the level of oxidative stress inside and outside cells. The fluorescence intensity of DCF is positively correlated with the ROS concentration. |
| 4-MUNANA
4-MUNANA is a substrate of influenza virus neuraminidase (NA) with high selectivity and irreversible reaction. In the enzymatic reaction, 4-MUNANA is hydrolyzed by NA to generate fluorescent 4-methylumbelliferone (4-MU). By detecting the fluorescence intensity of 4-MU, quantitative analysis of NA activity can be achieved. |
All AbMole products are for research use only, cannot be used for human consumption or veterinary use. We do not provide products or services to individuals. Please comply with the intended use and do not use AbMole products for any other purpose.
Products are for research use only. Not for human use. We do not sell to patients.
© Copyright 2010-2024 AbMole BioScience. All Rights Reserved.
