Cy5-DBCO (Synonyms: DBCO-Sulfo-Cy5) |
Catalog No.GC64831
One-Click Copy Product Info
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Cy5-DBCO is a bioorthogonal fluorescent probe that combines the near-infrared dye Cy5 with dibenzocyclooctyne (DBCO) (Ex=635nm, Em=650-700nm).
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 1564286-24-3
Sample solution is provided at 25 µL, 10mM.
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Cy5-DBCO is a bioorthogonal fluorescent probe that combines the near-infrared dye Cy5 with dibenzocyclooctyne (DBCO) (Ex=635nm, Em=650-700nm). Cy5-DBCO enables efficient and specific biomolecule labeling through a copper-free click chemistry reaction between the DBCO group and azides. Cy5-DBCO is used in applications such as bioimaging, protein labeling, cell tracing, and drug delivery system research[1-4].
References:
[1] Song S, Shim MK, Lim S, et al. In Situ One-Step Fluorescence Labeling Strategy of Exosomes via Bioorthogonal Click Chemistry for Real-Time Exosome Tracking In Vitro and In Vivo. Bioconjug Chem. 2020 May 20;31(5):1562-1574.
[2] Zhu L, Xu Y, Wei X, et al. Coupling Aptamer-based Protein Tagging with Metabolic Glycan Labeling for In Situ Visualization and Biological Function Study of Exosomal Protein-Specific Glycosylation. Angew Chem Int Ed Engl. 2021 Aug 9;60(33):18111-18115.
[3] Li L, Wang F, Mo S, et al. A Spatially Distributed Microneedle System for Bioorthogonal T Cell-Guided Cancer Therapy. Adv Sci (Weinh). 2025 Apr;12(13):e2416841.
[4] Xue Y, Li J, Chen D, et al. Observation of structural switch in nascent SAM-VI riboswitch during transcription at single-nucleotide and single-molecule resolution. Nat Commun. 2023 Apr 22;14(1):2320.
Method for Cy5-DBCO Cell Labeling and Tracing[1]
This protocol is adapted from research data and provided for reference only. Adjustments may be necessary based on specific experimental requirements.
1. Reagent Preparation: Dissolve tetraacetylated N-azidoacetyl-D-mannosamine (Ac4ManNAz) in complete cell culture medium to prepare a 50μM working solution. Dissolve Cy5-DBCO in a suitable buffer (DPBS, pH=7.4) to prepare a working concentration (20μM for in vitro experiments, 25μM for in vivo injection).
2. Target Cell Azide Labeling: Seed target cells at an appropriate density and culture them in complete medium containing 50μM Ac4ManNAz for 3 days to metabolically introduce azide groups onto the cell surface.
3. Cy5-DBCO Labeling and Detection (In Vitro): Wash the Ac4ManNAz-treated cells twice with DPBS (pH=7.4). Add the Cy5-DBCO working solution (20μM) and incubate at 37°C for 1 hour. Wash the cells thoroughly with DPBS to remove unbound probe. Subsequently, fix the cells (with formaldehyde-glutaraldehyde fixative), stain the nuclei (with DAPI), and perform fluorescence imaging analysis using a confocal laser scanning microscope (Ex=635nm, Em=650-700nm). Quantitative analysis can also be performed using flow cytometry.
4. Cy5-DBCO Labeling and Detection (In Vivo): Directly inject target cells treated with Ac4ManNAz for 3 days into the target organ of a live animal. Three days after cell transplantation, inject the Cy5-DBCO solution (25μM, 200μL) into the mouse via the tail vein. Four hours after probe injection, use an in vivo imaging system for whole-body fluorescence imaging to observe specific signals at the target organ. Alternatively, dissect the organ for ex vivo imaging using a fluorescence imaging station or prepare frozen sections for microscopic observation.
Precautions:
(1) Unreacted, free Cy5-DBCO is primarily cleared via the renal route within 4 hours post-injection, which helps reduce background signal.
(2) For your safety and health, please wear a lab coat and disposable gloves during operation.
References:
[1] Kang SW, Lee S, Na JH, et al. Cell labeling and tracking method without distorted signals by phagocytosis of macrophages. Theranostics. 2014 Feb 12;4(4):420-31.
| Cas No. | 1564286-24-3 | SDF | |
| Synonyms | DBCO-Sulfo-Cy5 | ||
| Formula | C52H56N4O11S3 | M.Wt | 1009.22 |
| Solubility | DMSO : 62.5 mg/mL (61.93 mM; Need ultrasonic); H2O : ≥ 10 mg/mL (9.91 mM) | 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. |
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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 | 990.9 μL | 4.9543 mL | 9.9086 mL |
| 5 mM | 198.2 μL | 990.9 μL | 1.9817 mL |
| 10 mM | 99.1 μL | 495.4 μL | 990.9 μL |
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 saline, 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: >97.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)