Physicochemical Properties
| Molecular Formula | C51H55CLN4O3 |
| Molecular Weight | 807.46 |
| Exact Mass | 806.396 |
| CAS # | 2270866-73-2 |
| PubChem CID | 162477420 |
| Appearance | Green to dark green solid powder |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 12 |
| Heavy Atom Count | 59 |
| Complexity | 1760 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | C1=CC=CC2=C3C(N(C)/C(=C\C=C4/CCCC(C=CC5=[N+](CCCCCC(NCCN6C(=O)C=CC6=O)=O)C6=CC=C7C=CC=CC7=C6C5(C)C)=C/4)/C3(C)C)=CC=C12.[Cl-] |t:17| |
| InChi Key | XCNYYRVWJSOPSW-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C51H54N4O3.ClH/c1-50(2)43(53(5)41-25-23-37-16-8-10-18-39(37)48(41)50)27-21-35-14-13-15-36(34-35)22-28-44-51(3,4)49-40-19-11-9-17-38(40)24-26-42(49)54(44)32-12-6-7-20-45(56)52-31-33-55-46(57)29-30-47(55)58;/h8-11,16-19,21-30,34H,6-7,12-15,20,31-33H2,1-5H3;1H |
| Chemical Name | 6-[(2Z)-1,1-dimethyl-2-[(2Z)-2-[3-[(E)-2-(1,1,3-trimethylbenzo[e]indol-3-ium-2-yl)ethenyl]cyclohex-2-en-1-ylidene]ethylidene]benzo[e]indol-3-yl]-N-[2-(2,5-dioxopyrrol-1-yl)ethyl]hexanamide;chloride |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| ln Vitro | The finest stock solution preparation 1. Making protein A 2 mg/mL protein (antibody) concentration is added to achieve the labeling effect. A pH of 8.5±0.5 is the ideal value for the protein solution. Use one milligram of carbon dioxide if the pH is less than 8.0. The labeling efficiency will be significantly decreased if the protein content is less than 2 mg/mL. Ultimately, a protein concentration range of 2–10 mg/mL is advised to achieve the optimal labeling efficiency. The labeling efficiency of the protein will be impacted if it is not placed in a transparent buffer that contains primary amines (like Tris or glycine) and ammonium ions. 2. Prepare the dye Establish a 10 mM stock solution by mixing CY dye with anhydrous DMSO. Before being aliquoted, the CY storage solution should be properly mixed using a glass tube or vortex and kept in a dark place at -20°C or -80°C. 3. The quantity of dye solution used Depending on how much protein has been labeled, CY dye will be needed for the labeling procedure. CY dye and protein should be taken at the following dosages: If 500 μL of 2 mg/mL IgG (MW = 150.000) is the necessary labeled protein, then 3.95 μL of CY volume is needed. To dissolve a tube containing 1 milligram of CY dye, use 100 μL of DMSO. To be more specific, the calculation procedure goes like this (using CY3-NHS ester as an example). mmol (IgG) = mg/mL (IgG) ×mL (IgG) / MW (IgG) = 2 mg/mL×0.5 mL / 150,000 mg/mmol = 6.7×10-6 mmol 2) mmol (CY3-NHS ester) = mmol (IgG) ×10 = 6.7×10-6 mmol×10 = 6.7×10-5 mmol Three) μL of CY3-NHS ester equals mmol of CY3-NHS ester, ×MW of CY3-NHS ester, / mg/μL of CY3-NHS ester, which translates to 6.7×10-5 mmol× 590.15 mg/mmol / 0.01 mg/μL, or 3.95 μL (by volume). Method of use 1. Reaction labeling 1) Slowly fill the fresh carrier with the calculated volume of 10 mg/mL CY dye. The centrifuged sample should be briefly collected at the bottom of the reaction tube. Add to the 0.5 mL protein sample solution, shake gently to mix. Never copy 2) Place the reaction tube in a dark area, give it a good shake, and walk for 60 minutes under the original settings (10–15 minutes each week). Gently flip the reaction tube over multiple times to reach 2. Protein desalting and blocking SepHadex G-25 column blocked dye conjugates are used as an example in the methodology that follows. Read the manufacturer's instructions and prepare the SepHadex G-25 column. 2) Fill the top of the SepHadex G-25 column with the reaction mixture. Upon reaching the lower limit of the upper resin's surface, incorporate PBS (pH 7.2-7.4). To complete columnar shrinking, apply extra PBS (pH 7.2–7.4) right away to the required sample. To create the desired dye-protein conjugate, combine the components. |
| References |
[1]. Biodegradable Zwitterionic Nanogels with Long Circulation for Antitumor Drug Delivery. ACS Appl Mater Interfaces. 2018 Jul 18;10(28):23509-23521. [2]. Ptaszek M. Rational design of fluorophores for in vivo applications. Prog Mol Biol Transl Sci. 2013;113:59-108. [3]. Shindy, H. A. (2017). Fundamentals in the chemistry of cyanine dyes: A review. Dyes and Pigments, 145, 505–513. doi:10.1016/j.dyepig.2017.06.029. |
Solubility Data
| 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 |
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.2385 mL | 6.1923 mL | 12.3845 mL | |
| 5 mM | 0.2477 mL | 1.2385 mL | 2.4769 mL | |
| 10 mM | 0.1238 mL | 0.6192 mL | 1.2385 mL |