Coppersensor-1 (CS1) is a membrane-permeable (penetrable) fluorescent dye. Coppersensor-1 has an affinity for Cu+ in the picomolar range and is selective for competing cellular metal ions. Coppersensor-1 serves as a probe that can selectively and sensitively detect copper(I) ions (Cu+) in biological samples including living cells. Coppersensor-1 can be used for imaging research into serious diseases such as cancer, cardiovascular disease, neurological diseases, and more.

Physicochemical Properties

Molecular Formula	C30H50BN3F2S4
Molecular Weight	629.806
Exact Mass	629.294
CAS #	874748-20-6
PubChem CID	101391795
Appearance	Brown to red solid powder
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	18
Heavy Atom Count	40
Complexity	877
Defined Atom Stereocenter Count	0
SMILES	[F-][B+3]1(N2=C(C(CC)=C(C)C2=C(CN(CCSCCSCC)CCSCCSCC)C2=C(C)C(CC)=C(C)[N-]12)C)[F-]
InChi Key	YPWFLOFNPLJPBT-UHFFFAOYSA-N
InChi Code	InChI=1S/C30H50BF2N3S4/c1-9-26-22(5)29-28(21-34(13-15-39-19-17-37-11-3)14-16-40-20-18-38-12-4)30-23(6)27(10-2)25(8)36(30)31(32,33)35(29)24(26)7/h9-21H2,1-8H3
Chemical Name	N-[(5,11-diethyl-2,2-difluoro-4,6,10,12-tetramethyl-3-aza-1-azonia-2-boranuidatricyclo[7.3.0.03,7]dodeca-1(12),4,6,8,10-pentaen-8-yl)methyl]-2-(2-ethylsulfanylethylsulfanyl)-N-[2-(2-ethylsulfanylethylsulfanyl)ethyl]ethanamine
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: This product requires protection from light (avoid light exposure) during transportation and storage.
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

Guidelines (The methodology listed below is our suggested one; it should be adjusted to meet your unique requirements as it just offers as a guideline)[1][2]. A. Making stock solutions of reagents for imaging: 1. Dissolve 0.63 mg of solid CS1 per milliliter of DMSO solvent to create a 1 mM stock solution (MW 630 g/moL) in DMSO. 2. Make a stock solution of CuCl or CuCl2 in 10 mM. B. Labeling living cells with CS1: 1. It will take 1-2 days for imaging investigations with cells in culture. Proceed with the regular protocol for incubating the cells. 2. Cells are passaged and plated on 18-mm glass coverslips coated with 50 μg/mL poly-L-lysine the day before imaging. For imaging, adherent cells are cultured to 50–80% confluency. 3. Take the cells out of the incubator and place one coverslip in a 35-mm Petri dish with three milliliters of PBS buffer in it. 4. To get a final dye concentration of 5 μM, add 15 μL of a 1 mM CS1 stock solution to the Petri dish. Stir well. (Higher dye concentrations may cause background fluorescence to be quite high.) 5. Incubate at 25 or 37°C for 5 to 20 minutes in the dark. C. Imaging the cells with CS1 label: 1. Imaging with a 543 nm excitation in a typical confocal microscope. (Any kind of fluorescence microscope, including confocal, multiphoton, and epifluorescence, can be used to image CS1.)

References

[1]. Preparation and use of Coppersensor-1, a synthetic fluorophore for live-cell copper imaging. Nat Protoc. 2006;1(2):824-7. [2]. A selective turn-on fluorescent sensor for imaging copper in living cells. J Am Chem Soc. 2006 Jan 11;128(1):10-1.

Solubility Data

Solubility (In Vitro)

DMSO: 25 mg/mL (39.69 mM)

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	1.5878 mL	7.9389 mL	15.8778 mL
	5 mM	0.3176 mL	1.5878 mL	3.1756 mL
	10 mM	0.1588 mL	0.7939 mL	1.5878 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.