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Filipin III

Catalog No.GC12048

Fluorescing antibiotic used for the detection of lipoproteins

Products are for research use only. Not for human use. We do not sell to patients.

Filipin III Chemical Structure

Cas No.: 480-49-9

Size Price Stock Qty
500ug
$155.00
In stock
1mg
$214.00
In stock
5mg
$850.00
In stock

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Sample solution is provided at 25 µL, 10mM.

Product has been cited by 3 publications

Product Documents

Quality Control & SDS

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Protocol

This plan only provides a guide, please modify it to meet your specific needs.

1. Preparation of cell membrane staining solution

(1) Prepare DMSO storage solution: Use DMSO or absolute ethanol to dissolve Filipin III and prepare a storage solution with a concentration of 1-5mM.

Note: Unused storage solution should be aliquoted and stored in the dark at -20°C or -80°C to avoid repeated freezing and thawing.

(2) Prepare working solution: Dilute the storage solution with an appropriate buffer (such as serum-free medium or PBS) and prepare a working solution with a concentration of 1-250 μM.

Note: Please adjust the concentration of the working fluid according to the actual situation and prepare it now.

 

2. Cell suspension staining

(1) Suspension cells: Centrifuge at 1000g for 3-5 minutes at 4°C, discard the supernatant, and wash twice with PBS for 5 minutes each time.

(2) Adherent cells: Wash twice with PBS, add trypsin to digest the cells, and centrifuge at 1000g for 3-5 minutes after digestion is completed.

(3) Add 1 mL of Filipin III working solution to resuspend the cells and incubate at room temperature in the dark for 15-30 minutes. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.

(4) After the incubation, centrifuge at 1000g for 5 minutes, remove the supernatant, add PBS and wash 2-3 times, 5 minutes each time.

(5) Resuspend the cells in pre-warmed serum-free cell culture medium or PBS. Observe by fluorescence microscopy or flow cytometry.

 

3. Cell adhesion staining

(1) Culture adherent cells on sterile coverslips.

(2) Remove the coverslip from the culture medium, suck out the excess culture medium, and place the coverslip in a humid environment.

(3) Add 100 μL of dye working solution from one corner of the coverslip and shake gently to evenly cover all cells with the dye.

(4) Incubate at room temperature in the dark for 15-30 minutes. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.

(5) After the incubation, discard the dye working solution and use pre-warmed culture solution to wash the coverslip 2 to 3 times.

4. Microscope detection: The excitation light range of Filipin III is 340-380nm, and the emission light range is 385-470 nm.

Precautions:

1) NoteFilipin fluorescent staining photobleaches very rapidly, thus the sample should be analyzed immediately.

2) For your safety and health, please wear a lab coat and disposable gloves.

Background

Cholesterol is both an important structural component of cell membranes and an early intermediate in hormone and bile acid biosynthesis. The localization and measurement of cholesterol in cells is therefore of great interest. Filipin is the collective name given to four isomeric polyene macrolides isolated from cultures of S. filipinensis; Filipin III is the predominant isomer and the one used in most studies. Filipin binds to cholesterol in membranes, forming ultrastructural aggregates and complexes which can be visualized by freeze-fracture electron microscopy.[1],[2] The binding of cholesterol also decreases the intrinsic fluorescence of Filipin, and this property has also been used to detect cholesterol in membrane fractions.[3]

Reference:
[1]. Castanho, M.A., Coutinho, A., and Prieto, M.J. Absorption and fluorescence spectra of polyene antibiotics in the presence of cholesterol J. Biol. Chem. 267(1), 204-209 (1992).
[2]. Miller, R.G. Mini Review. The use and abuse of filipin to localize cholesterol in membranes Cell Biol. Int. Rep. 8(7), 519-535 (1984).
[3]. Severs, N.J., and Robenek, H. Detection of microdomains in biomembranes. An appraisal of recent developments in freeze-fracture cytochemistry Biochimica et Biophysica Acta 737, 373-408 (1983)

 

Chemical Properties

Cas No. 480-49-9 SDF
Chemical Name (3R,4S,6S,8S,10R,12R,14R,16S,17E,19E,21E,23E,25E,27S,28R)-4,6,8,10,12,14,16,27-octahydroxy-3-((R)-1-hydroxyhexyl)-17,28-dimethyloxacyclooctacosa-17,19,21,23,25-pentaen-2-one
Canonical SMILES O[C@@H]1/C(C)=C/C=C/C=C/C=C/C=C/[C@H](O)[C@@H](C)OC([C@@]([C@H](O)CCCCC)([H])[C@@H](O)C[C@@H](O)C[C@@H](O)C[C@@H](O)C[C@@H](O)C[C@@H](O)C1)=O
Formula C35H58O11 M.Wt 654.83
Solubility 1mg/ml in Ethanol; 5mg/ml in DMSO; 10mg/ml in DMF Storage Store at -20°C, protect from light.
We recommend using the stock solution within 24 hours or it may result in reduced activity.
General tips Please select the appropriate solvent to prepare the stock solution according to the solubility of the product in different solvents; once the solution is prepared, please store it in separate packages to avoid product failure caused by repeated freezing and thawing.Storage method and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored at -20°C, please use it within 1 month.
To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time.
Shipping Condition Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.
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In vivo Formulation Calculator (Clear solution)

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.

Related Video

    Filipin III- GlpBio

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Average Rating: 5 ★★★★★ (Based on Reviews and 5 reference(s) in Google Scholar.)

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