DiD perchlorate (Synonyms: D 307, DiIC18(5), NK 3175) |
| Catalog No.GC30543 |
DiD perchlorate is a lipophilic fluorescent dye that can rapidly and stably integrate into phospholipid cell membranes and is widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 127274-91-3
Sample solution is provided at 25 µL, 10mM.
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DiD perchlorate is a lipophilic fluorescent dye that can rapidly and stably integrate into phospholipid cell membranes and is widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins[1]. DiD exhibits distinct red fluorescence, which facilitates multicolor imaging and flow cytometry analysis of live cells[2]. DiD perchlorate has been used to label plasma membranes and endocytic organelles in bovine aortic endothelial cells and rat hippocampal slices[3]. DiD has also been used to assess proliferation in prostate cancer cell lines by flow cytometry, where cell populations with high DiD expression were associated with lower proliferation[3]. DiD is non-cytotoxic and detectable after three weeks in subcutaneously implanted PC3 cells in vivo[4]. DiD can be excited by 633nm helium-neon (He-Ne) laser, has a longer excitation and emission wavelength than Dil, and is especially suitable for labeling cells and tissues with background fluorescence.
References:
[1]. Yumoto, K., Berry, J.E., Taichman, R.S., et al.A novel method for monitoring tumor proliferation in vivo using fluorescent dye DiDCytometry A.85(6)548-555(2014).
[2]. Dailey, M.E., and Waite, M.Confocal imaging of microglial cell dynamics in hippocampal slice culturesMethods18(2)222-230(1999).
[3]. Lin, C.P., Lynch, M.C., and Kochevar, I.E.Reactive oxidizing species produced near the plasma membrane induce apoptosis in bovine aorta endothelial cellsExp. Cell Res.259(2)351-359(2000).
[4]. Ribeiro, T., Raja, S., Rodrigues, A.S., et al.NIR and visible perylenediimide-silica nanoparticles for laser scanning bioimagingDyes Pigments110227-234(2014)
This protocol provides a guide only and should be modified according to your specific needs.
1. Preparation of cell membrane staining solution
(1) Prepare DMSO or EtOH storage solution: the storage solution is prepared with DMSO or EtOH, the concentration is 1~5mM. For example, take 25mg of DiD (Mw: 959.91g/mol) and dissolve it in 5.21ml of anhydrous DMSO, fully dissolve to obtain a 5mM stock solution.
Note: Store unused stock solutions in aliquots at -20°C, 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 needs to be optimized according to different cell lines and experimental systems. It is recommended to start from the recommended concentration and explore the optimal concentration in the range of 10 times.
2. Suspension Cell Staining
(1) The suspended cells were centrifuged at 4°C, 1000-1500rpm for 3-5 minutes, and the supernatant was discarded. Wash twice with PBS for 5 minutes each.
(2) Add 1 mL of DiD perchlorate working solution and incubate at room temperature for 5-30 minutes.
Note: Different cells have different optimal culture time, 20min can be used as the initial incubation time, and then optimized to ensure uniform labeling results.
(3) After the incubation, centrifuge at 1000-1500rpm for 5 minutes, remove the supernatant, add PBS to wash 2-3 times, 5 minutes each time.
(4) Resuspend the cells with pre-warmed 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 coverslip from the culture medium, suck out the excess medium, and place the coverslip 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. Different cells have different optimal culture time, 20min can be used as the initial incubation time, and then optimized to ensure uniform labeling results.
(5) Discard the dye working solution after incubation, and wash the coverslip 2-3 times with pre-warmed culture solution.
4. Microscopic examination: the excitation/emission light of DiD perchlorate is 650/670nm 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. | 127274-91-3 | SDF | |
| Synonyms | D 307, DiIC18(5), NK 3175 | ||
| Canonical SMILES | CCCCCCCCCCCCCCCCCCN1C2=C(C=CC=C2)C(C)(C)/C1=C\C=C\C=C\C3=[N+](C4=C(C=CC=C4)C3(C)C)CCCCCCCCCCCCCCCCCC.O=Cl(=O)([O-])=O | ||
| Formula | C61H99ClN2O4 | M.Wt | 959.9 |
| Solubility | DMSO: 50 mg/mL (52.09 mM) | Storage | Store at -20°C,protect from light, stored under nitrogen,unstable in solution, ready to use. |
| 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 | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.0418 mL | 5.2089 mL | 10.4178 mL |
| 5 mM | 0.2084 mL | 1.0418 mL | 2.0836 mL |
| 10 mM | 0.1042 mL | 0.5209 mL | 1.0418 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: >95.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 15 reference(s) in Google Scholar.)
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