DiO (Synonyms: DiOC18(3); 3,3'-Dioctadecyloxacarbocyanine perchlorate) |
| Katalog-Nr.GC19515 |
3,3′-Dioctadecyloxacarbocyaninperchlorat ist ein grÜn fluoreszierender lipophiler Tracer, der in Wasser schwach fluoresziert, aber stark fluoresziert und ziemlich photostabil ist, wenn er in Membranen eingebaut wird.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 34215-57-1
Sample solution is provided at 25 µL, 10mM.
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DiO (3,3′-Dioctadecyloxacarbocyanine perchlorate) is a lipophilic long-chain carbocyanine fluorescent dye that is widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins. 3,3′-Octadecyloxacarbocyanine perchlorate is a green fluorescent lipophilic tracer that is weakly fluorescent in water but highly fluorescent and comparable when incorporated into membranes Photostability. 3,3′-Octadecyloxacarbocyanine perchlorate is used as a fluorescent marker for living cells and as a lipophilic dye for labeling zebrafish embryos[1].
Long-chain carbocyanines include DiO, DiI, DiD, DiR, and the dialkylaminostyrene dye DiA for labeling membranes and other hydrophobic structures. DiO fluorochrome exhibits pronounced fluorescence, which facilitates multicolor imaging and flow cytometry analysis of living cells[2]. DiO can be used with standard FITC filters.
References:
[1]. Anat Samuel, et al. Six3 regulates optic nerve development via multiple mechanisms. 2016 Jan 29;6:20267. doi: 10.1038/srep20267.
[2]. Bhowmik BB, et al. Photophysical studies of 3,3' dioctadecyloxacarbocyanine dye in model biological membranes and different solvents. Chem Phys Lipids. 2001 Feb;109(2):175-83.
This protocol provides a guide only and should be modified according to your specific needs.
1. DiO cell membrane staining solution
(1). Prepare DMSO or EtOH storage solution: use DMSO or EtOH as the storage solution with a concentration of 1-5mM.
Note: Store the unused stock solution in aliquots at -20°C in the dark and avoid repeated freezing and thawing.
(2). Preparation of working solution: Dilute the stock solution with a suitable buffer (such as: serum-free medium, HBSS or PBS) to prepare a working solution with a concentration of 0.5-5 μM.
Note: The final concentration of the working solution is prepared according to the experiences of different cells and experiments. Optimum conditions can be found from more than ten times the recommended concentration.
2. Suspension Cell Staining
(1). Centrifuge the suspended cells at 1000g for 3-5 minutes at 4°C and discard the supernatant. Wash twice with PBS for 5 minutes each.
(2). Add 1mL of DiO working solution and incubate at room temperature for 5-30 minutes. The optimal incubation time is different for different cells.
(3). Centrifuge the stained cell test tube at 1000~1500 rpm for 5 minutes.
(4). Centrifuge at 400g for 3-4 minutes at 4°C and discard the supernatant. Pour off the supernatant and add PBS to wash twice, 5 minutes each time.
(5). Resuspend the cells with serum-free cell culture medium or PBS. Observed by fluorescence microscopy or flow cytometry.
3. Staining of Adhesive Cells
(1). Culture adherent cells on sterile coverslips.
(2). Remove the cover slip from the medium, suck out the excess medium, and place the cover slip in a humid environment.
(3). Add 100uL of dye working solution from one corner of the cover slip and shake gently to make the dye evenly cover all the cells.
(4). Incubate at room temperature for 5-30 minutes, and the optimal incubation time is different for different cells.
(5). Discard the dye working solution and wash the coverslip 2~3 times with the culture medium.
4. Microscopic examination: the excitation/emission light of DiO is 484/501nm respectively.
Note:
1) Fluorescent dyes have quenching problems, please try to avoid light to slow down fluorescence quenching.
2) For your safety and health, please wear a lab coat and disposable gloves for operation.
| Cas No. | 34215-57-1 | SDF | |
| Überlieferungen | DiOC18(3); 3,3'-Dioctadecyloxacarbocyanine perchlorate | ||
| Canonical SMILES | O=Cl(=O)([O-])=O.CCCCCCCCCCCCCCCCCCN1C(C=CC=C2)=C2O/C1=C/C=C/C(OC3=C4C=CC=C3)=[N+]4CCCCCCCCCCCCCCCCCC | ||
| Formula | C₅₃H₈₅ClN₂O₆ | M.Wt | 881.7 |
| Löslichkeit | DMF : 10 mg/mL (11.34 mM; Need ultrasonic); DMSO : 5 mg/mL (5.67 mM; ultrasonic and warming and heat to 60°C) | Storage | Store at 4°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. |
||
| 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 | 1.1342 mL | 5.6709 mL | 11.3417 mL |
| 5 mM | 226.8 μL | 1.1342 mL | 2.2683 mL |
| 10 mM | 113.4 μL | 567.1 μL | 1.1342 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 11 reference(s) in Google Scholar.)
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