C11 BODIPY 581/591 |
Catalog No.GC40165 |
C11-BODIPY581/591 est une sonde de ratio fluorescente de l'oxydation des lipides.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 217075-36-0
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >97.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
This plan only provides a guide, please modify it to meet your specific needs.
1. Preparation of staining solution
(1) Prepare storage solution: Dissolve C11 BODIPY 581/591 in DMSO and prepare a storage solution with a concentration of 1-10mM.
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 a suitable buffer (such as serum-free medium or PBS) and prepare a working solution with a concentration of 1-10μM.
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 C11 BODIPY 581/591 working solution to resuspend the cells, and incubate at 37 °C in the dark for 5-30 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 cells in pre-warmed serum-free cell culture medium or PBS and observe by fluorescence microscope 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 37 °C in the dark for 5-30 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 and emission light of the oxidized form of C11 BODIPY 581/591 are 460-495nm and 510-550nm respectively; the excitation light and emission light of the reduced form are 565-581nm and 585-591nm respectively.
Precautions:
1) Fluorescent dyes all have quenching problems. Please avoid light as much as possible to slow down fluorescence quenching.
2) For your safety and health, please wear a lab coat and disposable gloves.
C11-BODIPY581/591 est un analogue fluorescent d'acide gras sensible à l'oxydation, avec des propriétés fluorescentes dans la plage rouge du spectre visible (maximum d'émission à 595 nm), ce qui permet son application en microscopie de fluorescence. C11-BODIPY581/591 s'intègre facilement dans les membranes et fluoresce en rouge à l'état intact mais passe au vert lorsqu'il est oxydé par des radicaux libres. Cette caractéristique est très avantageuse car elle permet le ratio-imaging des activités oxydantes au niveau (sub)cellulaire. De plus, les propriétés fluorescentes de C11-BODIPY581/591 permettent l'utilisation de cette sonde dans le criblage rapide et moyen d'antioxydants dans les cellules vivantes et les membranes modèles selon une approche multi-puits / lecteur de fluorescence.[1][2].
Les longueurs d'onde d'excitation et d'émission maximales du fluorophore C11-BODIPY581/591 correspondaient respectivement à 581 et 591 nm. L'ajout de CumOOH/hémine, en tant qu'initiateur de l'oxydation des lipides, a déplacé les spectres d'excitation et d'émission vers des longueurs d'onde plus courtes correspondant à une fluorescence verte (pic d'excitation à 500 nm, émission à 510 nm). C11-BODIPY581/591 est également facilement oxydé par d'autres systèmes générateurs de radicaux hydroxy-, peroxy- et oxy-, tels que le peroxyde d'hydrogène/Fe2+ et le 2,2'-azobis. Cependant, cette sonde est relativement insensible au SIN-1, qui génère de l'oxyde nitrique et du superoxyde[3].
References:
[1]. Drummen GP, et al. C11-BODIPY581/591, an oxidation-sensitive fluorescent lipid peroxidation probe: (micro)spectroscopic characterization and validation of methodology. Free Radic Biol Med. 2002 Aug 15;33(4):473-90.
[2]. Partyka A, et al. Detection of lipid peroxidation in frozen-thawed avian spermatozoa using C11-BODIPY581/591. Theriogenology. 2011 Jun;75(9):1623-9.
[3]. Pap EH, et al. Ratio-fluorescence microscopy of lipid oxidation in living cells using C11-BODIPY581/591. FEBS Lett. 1999 Jun 25;453(3):278-82.
Cas No. | 217075-36-0 | SDF | |
Chemical Name | (T-4)-difluoro[5-[[5-[(1E,3E)-4-phenyl-1,3-butadien-1-yl]-2H-pyrrol-2-ylidene-κN]methyl]-1H-pyrrole-2-undecanoato(2-)-κN1]-borate(1-), monohydrogen | ||
Canonical SMILES | [F-][B+3]1([N]2=C(/C=C/C=C/C3=CC=CC=C3)C=CC2=CC4=CC=C(CCCCCCCCCCC([O-])=O)[N-]14)[F-].[H+] | ||
Formula | C30H34BF2N2O2 • H | M.Wt | 504.4 |
Solubility | 30mg/ml in DMSO,Slightly soluble in Methanol | Storage | Store at -20°C |
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. |
Prepare stock solution | |||
1 mg | 5 mg | 10 mg | |
1 mM | 1.9826 mL | 9.9128 mL | 19.8255 mL |
5 mM | 0.3965 mL | 1.9826 mL | 3.9651 mL |
10 mM | 0.1983 mL | 0.9913 mL | 1.9826 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
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.
Average Rating: 5
(Based on Reviews and 33 reference(s) in Google Scholar.)GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
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