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Cy7 DiC18 (Synonyms: DiR)

Catalog No.GC35773

Cy7 DiC18 (DiIC 18(7); DiR) is a lipophilic long-chain carbocyanine dye that produces near-infrared fluorescence and is widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins.

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Cy7 DiC18 Chemical Structure

Cas No.: 100068-60-8

Size Price Stock Qty
10mM (in 1mL DMSO)
$80.00
In stock
5mg
$45.00
In stock
10mg
$72.00
In stock

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Sample solution is provided at 25 µL, 10mM.

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 or EtOH storage solution: The storage solution is prepared with DMSO or EtOH, with a concentration of 1~5mM.

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

(2) Preparation of working solution: Dilute the storage solution with an appropriate 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 based on experience with 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 time.

(2) Add 1 mL of dye working solution and incubate at room temperature in the dark for 5-30 minutes.

Note: The optimal culture time for different cells is different and can be adjusted according to specific experimental needs.

(3) After the incubation, centrifuge at 1000-1500g for 5 minutes, remove the supernatant, add PBS and wash 2-3 times, 5 minutes each time.

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

 

3. Staining of adherent cells

(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 100uL 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 culture time for different cells is different.

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

 

4. Exosome staining

(1) Take an appropriate volume of Di dye stock solution and add it to the collected exosome filtration supernatant to reach a final concentration of 5 μM, and mix gently using a pipette;

(2) Incubate at room temperature in the dark for 30 minutes, turning gently every 5 minutes.

(3) Transfer the incubated sample to a centrifuge tube and centrifuge it in an ultracentrifuge at 100000g and 4°C for 30 minutes;

(4) After centrifugation, carefully tilt and discard the supernatant in a clean bench. Use a micropipette with a sterile tip to remove the liquid that forms in the test tube but does not drip. This is the marked outer layer. secretion body;

Note: During this step, the free dye disappears with the decanted supernatant, so the solution can be decanted, but always be careful not to lose the precipitate.

5. Microscope detection: The excitation/emission light of Cy7 DiC18 (DiR) is 750/780nm 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.

Background

Cy7 DiC18 (DiIC 18(7); DiR) is a lipophilic long-chain carbocyanine dye that produces near-infrared fluorescence and is widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins. The two long 18-carbon chains of DiR insert into the cell membrane, resulting in specific and stable cell staining with little dye transfer between cells. DiR exhibits pronounced infrared fluorescence, which facilitates multicolor imaging and flow cytometry analysis of living cells. DiR is of great significance in in vivo imaging or tracking. The emitted infrared light can efficiently pass through cells and tissues, and the background fluorescence level in the infrared range is very low[1].

References:

[1]. Shiyan Fu,Ruihao Yang,Junjie Ren,Jiahui Liu,LeiZhang,Zhigang Xu,Yuejun Kang,and Peng Xue.Catalytically Active CoFe204 Nanoflowers for Augmented Sonodynamic and Chemodynamic Combination Therapy with Elicitation of Robust Immune Response.ACS Nano Article ASARDOI: 10.1021/acsnano.1c03128.

Chemical Properties

Cas No. 100068-60-8 SDF
Synonyms DiR
Canonical SMILES CCCCCCCCCCCCCCCCCC[N+]1=C(/C=C/C=C/C=C/C=C2N(CCCCCCCCCCCCCCCCCC)C3=C(C=CC=C3)C/2(C)C)C(C)(C)C4=C1C=CC=C4.[I-]
Formula C63H101IN2 M.Wt 1013.39
Solubility Soluble in DMSO Storage 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.

Complete Stock Solution Preparation Table

Prepare stock solution
1 mg 5 mg 10 mg
1 mM 0.9868 mL 4.9339 mL 9.8679 mL
5 mM 0.1974 mL 0.9868 mL 1.9736 mL
10 mM 0.0987 mL 0.4934 mL 0.9868 mL
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**When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and MSDS / CoA (available online).

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In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg 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.)

% DMSO % % Tween 80 % ddH2O
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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.

Related Video

    Cell and cell membrane - GlpBio

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Average Rating: 5 ★★★★★ (Based on Reviews and 29 reference(s) in Google Scholar.)

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