| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Cy5 amine TFA does not bind to a biological target; it is a chemical labeling reagent. The primary amine (-NH2) group can react with carboxyl groups via carbodiimide-mediated (EDC/NHS) chemistry to form stable amide bonds, or with activated ester groups (e.g., NHS esters) for covalent conjugation to proteins, peptides, or other biomolecules. The Cy5 fluorophore provides far-red fluorescence for detection.
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| ln Vitro |
In vitro, Cy5 amine TFA is used to prepare Cy5-labeled compounds or polymers, which can be used for imaging cellular processes and drug trafficking. The primary amine group enables conjugation to molecules containing carboxyl groups using standard EDC/NHS coupling chemistry. Typical procedure: dissolve the carboxyl-containing molecule (1 micromol) in 200 microL MES buffer (pH 5.5-6.0). Add 2-5 equivalents of EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide) and NHS (N-hydroxysuccinimide) to activate the carboxyl group for 15-30 minutes. Add 1-2 equivalents of Cy5 amine TFA (dissolved in DMSO or water) and adjust pH to 7.2-7.5. Incubate at room temperature for 2-4 hours or overnight at 4degC. The labeled product is purified by dialysis or HPLC and used for imaging applications.
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| ln Vivo |
Cy5 amine TFA is not administered directly for in vivo activity. It is used to label targeting ligands (e.g., antibodies, peptides, nanoparticles) via amine-carboxyl conjugation chemistry. The Cy5-labeled conjugates can be administered to animal models for far-red fluorescence imaging. Cy5 offers deeper tissue penetration (up to several millimeters) compared to Cy3, but less than NIR dyes like Cy7. This product is suitable for in vivo imaging of subcutaneous tumors and biodistribution studies.
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| Enzyme Assay |
For in vitro labeling of carboxyl-containing molecules via EDC/NHS chemistry: (1) Dissolve 1 micromol of carboxylate-containing biomolecule (e.g., hyaluronic acid, fatty acid, or peptide with free -COOH) in 200 microL of 0.1 M MES buffer (pH 5.5-6.0). (2) Add 2-5 micromol EDC and 2-5 micromol NHS (dissolved in water). Mix and incubate at room temperature for 15-30 minutes to activate the carboxyl groups. (3) Add 1-2 micromol Cy5 amine TFA (dissolved in 20 microL DMSO or 50 microL water) to the activation mixture. (4) Adjust the pH to 7.2-7.5 by adding 50-100 microL of 0.2 M sodium phosphate buffer (pH 7.5). (5) Incubate at room temperature for 2-4 hours with gentle shaking, or overnight at 4degC. (6) Purify the labeled product by dialysis (appropriate MWCO) or size-exclusion chromatography (e.g., Sephadex G-25 column). (7) Determine the degree of labeling by measuring absorbance at 650 nm (Cy5) and 280 nm (protein, if applicable). For protein labeling with NHS ester forms of Cy5, the amine form is not directly reactive; instead, use Cy5 NHS ester for amine-reactive labeling.
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| Cell Assay |
For cell-based assays, Cy5-labeled compounds or nanoparticles are used for cellular uptake imaging. Cells (e.g., 1×10⁵ cells/well in 8-well chamber slides) are seeded and cultured overnight. The Cy5-labeled probe (1-20 microg/mL) is added to the culture medium and incubated for 1-6 hours at 37degC. Cells are washed with PBS (3×), fixed with 4% paraformaldehyde (if not live-cell imaging), counterstained with Hoechst 33342 or DAPI for nuclei, and imaged using a fluorescence microscope with Cy5 filter (Ex 640-660 nm, Em 660-680 nm). For live-cell imaging of drug uptake, cells can be imaged directly after washing. For quantitative uptake, cells can be trypsinized, resuspended in PBS, and analyzed by flow cytometry using the Cy5 channel (excitation 640 nm, emission 670 nm).
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| Animal Protocol |
For in vivo animal studies, Cy5-labeled conjugates (e.g., antibodies, nanoparticles) are typically administered via intravenous (tail vein) or intraperitoneal injection to mice (6-8 weeks old, 20-25 g). Dose: 50-100 microg of labeled protein per mouse in 100-200 microL PBS. In vivo fluorescence imaging is performed using an imaging system with excitation at 640-650 nm and emission at 670-700 nm at various time points (e.g., 1, 6, 12, 24, 48 hours post-injection). For subcutaneous tumor models, mice are anesthetized with isoflurane during imaging. At the terminal endpoint, major organs are excised for ex vivo imaging to quantify biodistribution. Tumor-to-background signal ratios are calculated by comparing tumor fluorescence signal to adjacent muscle or skin.
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| ADME/Pharmacokinetics |
Cy5 amine TFA has a molecular weight of 731.33 Da (for the TFA salt). The compound is supplied as a solid (brown to dark brown solid) and should be stored at -20degC, protected from light and moisture. Powder is stable for up to 3 years at -20degC and 2 years at 4degC. Stock solutions in DMSO (10-20 mM) should be stored at -80degC for up to 6 months or at -20degC for up to 1 month. Solubility: DMSO (soluble, typical concentration 10-50 mg/mL); water (moderately soluble due to TFA salt; may require sonication to dissolve). For in vivo administration, the labeled conjugate (not the free dye) is used; the free dye is not administered directly. For conjugation, Cy5 amine TFA is typically dissolved in DMSO or water. The TFA salt form improves water solubility compared to the free base.
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| Toxicity/Toxicokinetics |
Cy5 amine TFA is considered non-toxic at typical research concentrations (microM to low mM range). Standard laboratory safety practices should be followed: use of gloves, lab coat, and eye protection; avoid inhalation of dust. This product is for research use only and not for human diagnostic or therapeutic applications. No significant toxicity has been reported. For in vivo studies, standard safety monitoring (body weight, behavior) should be performed.
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| References | |
| Additional Infomation |
Cy5 amine TFA (Cyanine5 amine TFA) is a fluorescent labeling dye for the far-red/NIR region. Key features: (1) Cy5 fluorophore with excitation/emission maxima at approximately 650/670 nm (typical) or 645/665 nm; (2) Primary amine functional group (-NH2) enables conjugation to carboxyl groups via EDC/NHS chemistry; (3) TFA salt form enhances water solubility and stability; (4) Far-red emission provides low background autofluorescence from biological samples, suitable for in vitro and in vivo imaging. Common applications: labeling of nanoparticles, polymers, peptides, and small molecules for cellular uptake, drug trafficking, and biodistribution studies. The compound is also used to prepare Cy5.5-labeled compounds (Cy5.5 is a derivative with slightly red-shifted spectra). Purity is typically ≥98% by HPLC. Related CAS for free base (Cy5 amine): 1807529-70-9.
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| Molecular Formula |
C40H54CLF3N4O3
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|---|---|
| Molecular Weight |
731.33
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| Related CAS # |
Cy 5 amine;1807529-70-9
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| Appearance |
Brown to dark brown solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3674 mL | 6.8369 mL | 13.6737 mL | |
| 5 mM | 0.2735 mL | 1.3674 mL | 2.7347 mL | |
| 10 mM | 0.1367 mL | 0.6837 mL | 1.3674 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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