3,3'-Dihexyloxacarbocyanine iodide (DiOC6(3) iodide) (Synonyms: DiOC6(3), NK 2280) |
| カタログ番号GC33493 |
3,3'-ヨウ化ジヘキシルオキサカルボシアニン (DiOC6(3) ヨウ化物) は、ミトコンドリア膜電位の変化を監視するために使用できるカルボシアニン色素です。
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 53213-82-4
Sample solution is provided at 25 µL, 10mM.
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3,3'-Dihexyloxacarbocyanine iodide is a carbocyanine dye which can be used to monitor changes in mitochondrial membrane potential.
Addition of cells to the cuvette containing 0.25 μM 3,3'-Dihexyloxacarbocyanine iodide (DiOCg(3)) leads to an increase in fluorescence, equilibration is rapid and is complete by 4 min. When f-met-leu-phe (10-7 M) is added to neutrophils preequilibrated with 3,3'-Dihexyloxacarbocyanine iodide, there is a short lag period of 5 seconds. The lag period is followed by a rapid loss of fluorescence. Examination of 3,3'-Dihexyloxacarbocyanine iodide loaded neutrophils using fluorescence microscopy demonstrates that in resting neutrophils the brightest fluorescence is associated with long thin organelles which corresponds to the distribution of mitochondria[1].
[1]. Korchak HM, et al. A carbocyanine dye, DiOC6(3), acts as a mitochondrial probe in human neutrophils. Biochem Biophys Res Commun. 1982 Oct 29;108(4):1495-501.
This plan only provides a guide, please modify it to meet your specific needs.
1. Preparation of staining solution
(1) Prepare DMSO stock solution: Dissolve DiOC6(3) iodide in DMSO and prepare a stock solution with a concentration of 1~5mM.
Note: Unused stock solution should be aliquoted and stored in the dark at -20°C to avoid repeated freezing and thawing.
(2) Prepare working solution: Dilute the stock solution with a suitable buffer (such as serum-free medium or PBS) and prepare a working solution with a concentration of 0.5~5 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 1mL of DiOC6 (3) iodide working solution and resuspend about 106 cells. Incubate at room temperature in 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 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 100μL 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 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 maximum excitation/emission light of DiOC6(3) iodide is 484/501 nm 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.
| Cas No. | 53213-82-4 | SDF | |
| 同義語 | DiOC6(3), NK 2280 | ||
| Canonical SMILES | CCCCCCN1/C(OC2=CC=CC=C12)=C/C=C/C3=[N+](CCCCCC)C4=CC=CC=C4O3.[I-] | ||
| Formula | C29H37IN2O2 | M.Wt | 572.52 |
| 溶解度 | DMSO : ≥ 125 mg/mL (218.33 mM);Water : < 0.1 mg/mL (insoluble) | Storage | Store at 2-8°C, protect from light |
| 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. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.7467 mL | 8.7333 mL | 17.4666 mL |
| 5 mM | 0.3493 mL | 1.7467 mL | 3.4933 mL |
| 10 mM | 0.1747 mL | 0.8733 mL | 1.7467 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
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.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 34 reference(s) in Google Scholar.)
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