| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
Sulfo-Cy7.5 NHS ester is not a pharmaceutical drug and does not bind to a biological receptor; it is a chemical labeling reagent. Its target is primary amine groups (-NH2) present on proteins, peptides, and antibodies. Upon conjugation, the fluorescent dye becomes covalently attached to the target molecule, enabling its visualization and tracking in biological systems. The sulfonate groups confer high water solubility, which reduces nonspecific binding and improves handling in aqueous buffers.
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| ln Vitro |
In vitro, Sulfo-Cy7.5 NHS ester is used to fluorescently label purified proteins, antibodies, and peptides. The NHS ester group is highly reactive with primary amines under mild conditions, forming stable amide bonds. The near-infrared fluorescence (Ex/Em 778/797 nm) allows for deep tissue penetration and minimal background autofluorescence, making it particularly useful for live-cell imaging and in vivo imaging studies. The labeled biomolecules retain their biological activity and are suitable for various applications, including Western blotting, ELISA, flow cytometry, and fluorescence microscopy.
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| ln Vivo |
In vivo, Sulfo-Cy7.5 NHS ester is not administered directly but is used to pre-label biomolecules (e.g., antibodies, peptides, nanoparticles) that are then administered to animals for near-infrared fluorescence imaging studies. The sulfo groups ensure good water solubility and reduce aggregation, while the NIR fluorophore enables non-invasive imaging of tissue distribution, tumor targeting, and biodistribution in living mice. The long wavelength minimizes light scattering and absorption by hemoglobin and water, allowing imaging depths of up to 1-2 cm. This is valuable for tracking drug delivery, visualizing tumor margins, and assessing therapeutic efficacy in preclinical models.
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| Enzyme Assay |
For non-cell-based labeling assays, a standard protocol is used to conjugate Sulfo-Cy7.5 NHS ester to a protein of interest. The protein (e.g., IgG antibody, 2-10 mg/mL) is prepared in PBS (pH 7.4) or 0.1 M sodium bicarbonate buffer (pH 8.5). The dye is dissolved in anhydrous DMSO to a concentration of 10 mM. A 10- to 20-fold molar excess of dye is added to the protein solution and mixed gently. The reaction is incubated at room temperature for 60 minutes or at 4degC overnight, protected from light. Unconjugated dye is removed by size exclusion chromatography (e.g., Sephadex G-25) or dialysis. The degree of labeling (DOL) is calculated by measuring absorbance at 280 nm (protein) and 778 nm (dye) using a spectrophotometer.
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| Cell Assay |
For in vitro cell assays, cells are seeded in glass-bottom culture dishes and allowed to adhere overnight. A pre-labeled protein (e.g., Sulfo-Cy7.5-labeled anti-EGFR antibody, 10 ug/mL) is added to the culture medium and incubated at 37degC for 30-60 minutes. For live-cell imaging, cells are washed three times with PBS to remove unbound labeled antibody and maintained in phenol-red-free medium. Fluorescence images are acquired using a confocal microscope equipped with a 785 nm laser and 800-830 nm emission filter. For fixed-cell studies, cells are fixed with 4% paraformaldehyde before imaging. The signal-to-noise ratio is typically high due to minimal background autofluorescence in the NIR range.
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| Animal Protocol |
For in vivo animal studies, an antibody-dye conjugate prepared using Sulfo-Cy7.5 NHS ester is administered intravenously to tumor-bearing mice (e.g., nude mice with subcutaneous tumor xenografts expressing a target antigen). Mice receive 50-200 ug of labeled antibody (concentration 0.5-2 mg/mL) via tail vein injection. Anesthesia is induced with 2% isoflurane, and NIR fluorescence imaging is performed at various time points (1, 6, 12, 24, 48, 72 hours post-injection) using an in vivo imaging system with 745 nm excitation and 800 nm emission filters. The biodistribution of the labeled antibody is assessed by measuring fluorescence intensity in the tumor and major organs (liver, kidney, spleen, lung, heart).
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| ADME/Pharmacokinetics |
Sulfo-Cy7.5 NHS ester is a water-soluble near-infrared dye with excitation maximum at 778 nm and emission maximum at 797 nm. It has a high extinction coefficient of 222,000 M-¹cm-¹ and a fluorescence quantum yield of 0.21. The NHS ester group is prone to hydrolysis in aqueous solution, so the dye should be dissolved in DMSO immediately before use and added to protein solutions within 1-2 hours for optimal conjugation efficiency. The labeled protein should be stored at 4degC or -80degC after adding a preservative such as glycerol (50%) for long-term storage.
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| Toxicity/Toxicokinetics |
The NHS ester group is chemically reactive and can cause skin and eye irritation upon direct contact. Standard chemical safety practices should be followed: wear gloves, a lab coat, and safety goggles when handling this compound. Avoid inhalation of dust. In solution, the dye should be protected from light to prevent photobleaching. Hydrolysis of the NHS ester generates N-hydroxysuccinimide, which is of low toxicity. This compound is not intended for human therapeutic or diagnostic use and should be treated as a research chemical. No specific acute toxicity data is available.
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| References | |
| Additional Infomation |
Sulfo-Cy7.5 NHS ester is a popular choice for in vivo imaging and flow cytometry due to its excellent water solubility and NIR fluorescence properties, which minimize tissue autofluorescence and allow deep tissue penetration. The trisulfonated structure prevents aggregation in biological fluids and reduces nonspecific binding to cell membranes, improving the signal-to-noise ratio in imaging experiments. This dye is commonly used for fluorescence-guided surgery, tumor imaging, and evaluation of drug delivery systems in preclinical research. The product is for research use only and is not for clinical diagnostics or therapy.
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| Molecular Formula |
C49H52KN3O16S4
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|---|---|
| Molecular Weight |
1106.30599021912
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| Exact Mass |
1103.171
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| CAS # |
2736437-44-6
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| PubChem CID |
171975657
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| Appearance |
Light green to green solid powder
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
17
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| Rotatable Bond Count |
13
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| Heavy Atom Count |
73
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| Complexity |
2770
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1(C(=[N+](CCCCCC(=O)ON2C(CCC2=O)=O)C2C=CC3=C(C=C(S([O-])(=O)=O)C=C3C1=2)S(O)(=O)=O)C=CC1CCCC(=CC=C2N(C3C=CC4=C(C=C(S(O)(=O)=O)C=C4C=3C2(C)C)S(O)(=O)=O)C)C=1)C.[KH]
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| InChi Key |
HDBJLDCQQJVKRA-UHFFFAOYSA-M
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| InChi Code |
InChI=1S/C49H51N3O16S4.K/c1-48(2)41(50(5)37-17-15-33-35(46(37)48)25-31(69(56,57)58)27-39(33)71(62,63)64)19-13-29-10-9-11-30(24-29)14-20-42-49(3,4)47-36-26-32(70(59,60)61)28-40(72(65,66)67)34(36)16-18-38(47)51(42)23-8-6-7-12-45(55)68-52-43(53)21-22-44(52)54;/h13-20,24-28H,6-12,21-23H2,1-5H3,(H3-,56,57,58,59,60,61,62,63,64,65,66,67);/q;+1/p-1
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| Chemical Name |
potassium;(2Z)-3-[6-(2,5-dioxopyrrolidin-1-yl)oxy-6-oxohexyl]-1,1-dimethyl-2-[(2Z)-2-[3-[(E)-2-(1,1,3-trimethyl-6,8-disulfobenzo[e]indol-3-ium-2-yl)ethenyl]cyclohex-2-en-1-ylidene]ethylidene]benzo[e]indole-6,8-disulfonate
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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: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
DMSO : ~50 mg/mL (~45.28 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.25 mg/mL (1.13 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1.25 mg/mL (1.13 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.9039 mL | 4.5195 mL | 9.0391 mL | |
| 5 mM | 0.1808 mL | 0.9039 mL | 1.8078 mL | |
| 10 mM | 0.0904 mL | 0.4520 mL | 0.9039 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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