| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
Not applicable; this is a fluorescent labeling dye, not a biologically active drug.
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|---|---|
| ln Vitro |
No direct biological activity is attributed to Cyanine5.5 NHS ester. In vitro, the NHS ester reacts with primary amines on proteins, forming stable amide bonds. The labeled proteins (e.g., antibodies) are then used for immunofluorescence, Western blotting, or flow cytometry. The dye can also intercalate into double-helical DNA, exhibiting enhanced fluorescence upon binding. However, this intercalation is not sequence-specific and is typically not used as a drug mechanism. The dye does not inhibit enzymes or modulate signaling pathways. No classical pharmacological activity is measured for the free dye.
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| ln Vivo |
No direct in vivo activity is attributed to the free dye. Cyanine5.5 NHS ester can be conjugated to antibodies, peptides, or nanoparticles for in vivo near-infrared fluorescence imaging in animal models. The conjugate is administered intravenously to mice. The far-red fluorescence (Ex/Em 675/694 nm) allows imaging of deep tissues with reduced autofluorescence. The dye does not have any intrinsic therapeutic or pharmacodynamic activity. No in vivo efficacy studies are conducted with the unconjugated dye. The NHS ester is not stable in aqueous buffers for extended periods; it must be conjugated immediately before use or stored in anhydrous DMSO.
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| Enzyme Assay |
The free dye does not bind to any specific receptor or enzyme. Its NHS ester reacts chemically with amine groups but does not engage in biological target recognition. For quality control, the absorption spectrum is measured in methanol or DMSO to confirm the peak at 675 nm. The dye can be analyzed by HPLC-MS. The NHS ester can be characterized by FT-IR (presence of a band at ∼1740 cm-¹). No classical receptor binding assays (radioligand displacement, enzyme inhibition) are performed for this compound, as it is a labeling reagent.
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| Cell Assay |
The dye is not used directly on cells; it must first be conjugated to a targeting molecule. For conjugation, dissolve Cyanine5.5 NHS ester in anhydrous DMSO (10 mM). Add the dye solution to a protein (e.g., antibody at 1 mg/mL in PBS, pH 7.4, or 0.1 M sodium bicarbonate buffer, pH 8.5) at a 10-20 fold molar excess. Incubate at room temperature for 1 hour with gentle shaking. Protect from light. Remove unreacted dye by dialysis, size-exclusion chromatography, or spin filtration. For cell staining, add the labeled protein (e.g., 1-20 ug/mL) to cultured cells for 30-60 min at 37degC or 4degC. Wash twice with PBS. Image by fluorescence microscopy using a Cy5.5 filter set (excitation 665-685 nm, emission 690-710 nm) or analyze by flow cytometry.
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| Animal Protocol |
The dye is not administered directly; it is first conjugated to a targeting vector. For imaging studies, conjugate Cyanine5.5 NHS ester to an antibody or peptide using the protocol above. After purification, the conjugate is administered intravenously (tail vein) to mice at a dose of 0.5-5 mg/kg (based on conjugate). At time points of 1, 6, 24, 48, and 72 hours post-injection, anesthetize the mice and obtain whole-body fluorescence images using an IVIS system equipped with a Cy5.5 filter set (excitation 660-680 nm, emission 690-710 nm). Quantify fluorescence intensity in tumors, major organs, and blood. Euthanize mice and collect organs for ex vivo imaging. All studies require IACUC approval. The unconjugated dye is not used in animals.
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| ADME/Pharmacokinetics |
No formal PK data exists for the free dye. The free NHS ester is unstable in aqueous solutions; its half-life in PBS (pH 7.4) at room temperature is approximately 1 hour. For the conjugated form (dye-protein), the PK is dictated by the carrier (e.g., antibody). Solubility: Soluble in DMSO (≥10 mM) and DMF. Store powder at -20degC in a desiccated, light-protected container. Stock solutions in anhydrous DMSO can be stored at -80degC for up to 6 months. Protect from light and moisture. The NHS ester is moisture-sensitive; vials should be allowed to warm to room temperature before opening to prevent condensation.
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| Toxicity/Toxicokinetics |
No specific toxicology data is available. As an NHS ester, the compound is an irritant and can cause skin, eye, and respiratory tract irritation. The compound is also moisture-sensitive and may release heat upon hydrolysis. Standard laboratory safety precautions must be followed: use in a fume hood, wear nitrile gloves, lab coat, and safety goggles. Avoid dust and aerosol formation. The compound is not approved for human or veterinary use. It is strictly for research use only. In animal imaging studies at typical doses (<5 mg/kg of conjugate), no overt toxicity has been reported. The free dye should not be injected.
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| References | |
| Additional Infomation |
Cyanine5.5 NHS ester tetrafluoroborate is a far-red/near-infrared fluorescent dye used for labeling primary amines on proteins, antibodies, and other biomolecules. Cy5.5 has excitation/emission maxima of 675/694 nm, making it ideal for in vivo imaging due to deep tissue penetration and low autofluorescence. The NHS ester reacts rapidly with amines under mild conditions to form stable amide bonds. In addition to protein labeling, the dye can intercalate into double-helical DNA, with enhanced fluorescence upon binding. Common applications include in vivo tumor imaging, flow cytometry, and fluorescence microscopy. This product is not a drug and has not been approved for clinical or diagnostic use. It is strictly a research chemical.
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| Molecular Formula |
C44H46BF4N3O4
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|---|---|
| Molecular Weight |
767.66
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| Exact Mass |
767.351
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| CAS # |
2375105-86-3
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| PubChem CID |
164577385
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| Appearance |
Typically exists as solid at room temperature
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
11
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| Heavy Atom Count |
56
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| Complexity |
1470
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| Defined Atom Stereocenter Count |
0
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| SMILES |
F[B-](F)(F)F.O(C(C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])N1C2C([H])=C([H])C3=C([H])C([H])=C([H])C([H])=C3C=2C(C([H])([H])[H])(C([H])([H])[H])/C/1=C(/[H])\C(\[H])=C(/[H])\C(\[H])=C(/[H])\C1C(C([H])([H])[H])(C([H])([H])[H])C2C3=C([H])C([H])=C([H])C([H])=C3C([H])=C([H])C=2[N+]=1C([H])([H])[H])=O)N1C(C([H])([H])C([H])([H])C1=O)=O
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| InChi Key |
QJHUOARMFFEDJW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C44H46N3O4.BF4/c1-43(2)36(45(5)34-25-23-30-16-11-13-18-32(30)41(34)43)20-8-6-9-21-37-44(3,4)42-33-19-14-12-17-31(33)24-26-35(42)46(37)29-15-7-10-22-40(50)51-47-38(48)27-28-39(47)49;2-1(3,4)5/h6,8-9,11-14,16-21,23-26H,7,10,15,22,27-29H2,1-5H3;/q+1;-1
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| Chemical Name |
(2,5-dioxopyrrolidin-1-yl) 6-[1,1-dimethyl-2-[5-(1,1,3-trimethylbenzo[e]indol-3-ium-2-yl)penta-2,4-dienylidene]benzo[e]indol-3-yl]hexanoate;tetrafluoroborate
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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 |
| 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 | 1.3027 mL | 6.5133 mL | 13.0266 mL | |
| 5 mM | 0.2605 mL | 1.3027 mL | 2.6053 mL | |
| 10 mM | 0.1303 mL | 0.6513 mL | 1.3027 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.
Products are for research use only; We do not sell to patients
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