NEP (VDP-green (NEP)) is a sensing fluorescent probe based on the intramolecular charge transfer (ICT) mechanism for sensing ortho-dithiol-containing proteins (VDPs). NEP displays high selectivity for VDPs in living cells and animals and displays strong green fluorescence signal (λex/λem=430/535 nm). NEP may be utilized in PD/Parkinson's disease study.

Physicochemical Properties

Molecular Formula	C29H31ASN4O5S2
Molecular Weight	654.63
Exact Mass	654.095
CAS #	2414276-32-5
PubChem CID	162642078
Appearance	Typically exists as solid at room temperature
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	11
Heavy Atom Count	41
Complexity	968
Defined Atom Stereocenter Count	0
InChi Key	GNUHSLLRVGIPIB-UHFFFAOYSA-N
InChi Code	InChI=1S/C29H31AsN4O5S2/c1-2-3-15-34-27(36)22-6-4-5-21-24(12-11-23(26(21)22)28(34)37)33-29(38)39-16-20-17-40-30(41-20)18-7-9-19(10-8-18)32-25(35)13-14-31/h4-12,20H,2-3,13-17,31H2,1H3,(H,32,35)(H,33,38)
Chemical Name	[2-[4-(3-aminopropanoylamino)phenyl]-1,3,2-dithiarsolan-4-yl]methyl N-(2-butyl-1,3-dioxobenzo[de]isoquinolin-6-yl)carbamate
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	HepG2 and PC12 cells show negligible or no cytotoxicity when exposed to NEP (1-30 μM; 6 hours)[1]. The fluorescence intensity of PC12 cells pretreated with 6-OHDA (0, 50, 100, and 200 μM; for 30 min) gradually decreases when exposed to NEP (10 μM; 4 hours)[1]. As the VDP receptor, NEP has a dithiarsolane moiety, which is a five-membered As-S ring. When VDPs are present, NEP exhibits a robust green fluorescence signal that is caused by the intramolecular cyclization that follows the cyclic dithiarsolane cleavage, which releases the fluorophore. NEP sustains a consistent fluorescence response between pH 7 and pH 10[1].
ln Vivo	Zebrafish that are 4 days old exhibit a clear green signal when exposed to 10 μM NEP for 4 hours[1].
References	[1]. Decrease of Protein Vicinal Dithiols in Parkinsonism Disclosed by a Monoarsenical Fluorescent Probe. Anal Chem. 2020 Mar 17;92(6):4371-4378.

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	1.5276 mL	7.6379 mL	15.2758 mL
	5 mM	0.3055 mL	1.5276 mL	3.0552 mL
	10 mM	0.1528 mL	0.7638 mL	1.5276 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.