To address white precipitations observed during Filipin staining for endothelial cells, increasing the amount of DMSO in the final staining solution may be necessary. This can be achieved by diluting the 1 mg/ml Filipin III solution with more DMSO solution to create a 0.5 mg/ml or 0.1 mg/ml concentrated solution prior to staining the samples.
For improving cholesterol content measurement in HUVEC cells, optimize the working concentration of Filipin and conduct a time-course experiment to enhance staining results. Prompt analysis of stained samples can help prevent photobleaching.
SAE0087
Filipin III ready made solution
from Streptomyces filipinensis, 1mg/ml in DMSO based solution
Synonym(s):
Filipin III solution
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About This Item
UNSPSC Code:
12352200
NACRES:
NA.77
Pricing and availability is not currently available.
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Quality Level
Assay
≥95% (HPLC)
form
ready-to-use solution
storage condition
protect from light
shipped in
dry ice
storage temp.
−20°C
Related Categories
General description
Filipin III is a polyene macrolide antibiotic and the major component isolated from cultures of S. filipinensis. 1 Due to its ability to fluoresce and bind to cholesterol, Filipin has been widely
used as a probe for sterol location in biological membranes 2 and has been used clinically as a stain for free cholesterol in the study of Type C Niemann-Pick disease. 3 Filipin inhibits prion protein (PrP) endocytosis and causes the release of PrP from the plasma membrane. 4 Filipin III was found to trigger signaling responses in tobacco cells, including a NADPH oxidase-dependent production. 5. The antifungal mechanism of action may be due to altering membrane permeability and associated functions by binding to membrane sterols. Filipin inhibits prion protein (PrP) endocytosis and causes the release of PrP from the plasma membrane.
used as a probe for sterol location in biological membranes 2 and has been used clinically as a stain for free cholesterol in the study of Type C Niemann-Pick disease. 3 Filipin inhibits prion protein (PrP) endocytosis and causes the release of PrP from the plasma membrane. 4 Filipin III was found to trigger signaling responses in tobacco cells, including a NADPH oxidase-dependent production. 5. The antifungal mechanism of action may be due to altering membrane permeability and associated functions by binding to membrane sterols. Filipin inhibits prion protein (PrP) endocytosis and causes the release of PrP from the plasma membrane.
Application
Filipin has been used in a double staining procedure as a probe for the detection of lipoproteins in polyacrylamide gel and immobilized on nitrocellulose membranes. It is also widely used to localize and quantitate unesterified cholesterol by virtue of a specific fluorescent complex.
Other Notes
- Filipin III solution is very sensitive to air and light. Upon receipt aliquot and store at -20°C avoid freeze thaw cycles.
- Filipin III is used in various concentrations depending on the specific protocol used. Dilute 1mg/ml stock solution with appropriate buffer according to the used protocol.
- Filipin III interaction with cholesterol alters the absorption and fluorescence spectra, for visualization with a fluorescence microscope use excitation at 340-380 nm and emission at 385-470 nm (Filipin fluorescent staining photo-bleaches very rapidly, thus the sample should be analyzed immediately).
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
188.6 °F
Flash Point(C)
87 °C
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The alterations in the absorption and fluorescence spectra observed for the polyene antibiotics filipin and nystatin in the presence of cholesterol are due to an exciton interaction (polyene aggregates) and cannot be attributed to a specific sterol-antibiotic complex. Filipin and
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Any suggestions for addressing white precipitations observed during Filipin staining for endothelial cells, possibly due to DMSO? Also, seeking advice to improve the experiment where most cells were negative for Filipin staining, and only a few were positive while measuring cholesterol content in HUVEC cells.
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