IR 813 perchlorate is a near-infrared (NIR) fluorescent dye (Ex=815 nm, Em=840 nm) that may be utilized to visualize regional lymph nodes in mice.

Physicochemical Properties

Molecular Formula	C40H40N2CL+.O4CL-
Molecular Weight	683.6626
Exact Mass	632.221
CAS #	201024-57-9
Related CAS #	IR 813 tosylate;134127-48-3
PubChem CID	6508807
Appearance	Typically exists as solid at room temperature
Melting Point	238-242ºC
LogP	9.554
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	3
Heavy Atom Count	48
Complexity	1260
Defined Atom Stereocenter Count	0
SMILES	O=Cl([O-])(=O)=O.ClC1/C(=C/C=C2/N(C)C3C=CC4=CC=CC=C4C=3C/2(C)C)/CCCC=1C=CC1=[N+](C)C2C=CC3=CC=CC=C3C=2C1(C)C |t:33|
InChi Key	QUUNBGLRLXAGFO-UHFFFAOYSA-M
InChi Code	InChI=1S/C40H40ClN2.ClHO4/c1-39(2)34(42(5)32-22-18-26-12-7-9-16-30(26)36(32)39)24-20-28-14-11-15-29(38(28)41)21-25-35-40(3,4)37-31-17-10-8-13-27(31)19-23-33(37)43(35)6;2-1(3,4)5/h7-10,12-13,16-25H,11,14-15H2,1-6H3;(H,2,3,4,5)/q+1;/p-1
Chemical Name	(2Z)-2-[(2E)-2-[2-chloro-3-[(E)-2-(1,1,3-trimethylbenzo[e]indol-3-ium-2-yl)ethenyl]cyclohex-2-en-1-ylidene]ethylidene]-1,1,3-trimethylbenzo[e]indole;perchlorate
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
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	IR 813 (17.3 μM) induces red blood cell hemolysis at a rate of 31.4% [1]. In MRC-5 cells, IR 813 (5.9 μM) causes 50% cell death within 24 hours [1].
ln Vivo	IR 813 perchlorate demonstrated fast extravasation and peak fluorescence intensity four hours after injection when used to monitor regional lymph nodes in mice [1]. Advice (This is our suggested protocol, which should be adjusted to suit your particular requirements as it only offers guidance) [1]. 1. Animals: In order to decrease tissue autofluorescence in the NIR region, ten-week-old female Balb/cOlaAnN mice were given an unlimited supply of food and water and kept on a 12-hour light/dark cycle. 2. Dosage: The mouse was given a single subcutaneous injection in its right front paw of 5.1 nmol IR 813 dye (20 μL 0.173 mg/mL, dissolved in PEG-400/ethanol/water=3:2:5, v/v/v solution). ..3. For in vivo optical imaging, utilize the Fluobeam700 NIR imaging system (739 nm excitation light, 3.5 mW), with a 750 nm long-pass emission cutoff filter. A week's worth of data on the fluorescence intensity in axillary lymph nodes (ALN) was collected (5 minutes, 1 hour, 4 hours, 24 hours, and 7 days). 4. Data analysis: To extract semi-quantitative information from fluorescence images, use ImageJ 1.44 software.
References	[1]. Surface chemistry architecture of silica nanoparticles determine the efficiency of in vivo fluorescence lymph node mapping. ACS Nano. 2013 Oct 22;7(10):8645-57.

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	1.4627 mL	7.3136 mL	14.6272 mL
	5 mM	0.2925 mL	1.4627 mL	2.9254 mL
	10 mM	0.1463 mL	0.7314 mL	1.4627 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.