L 012 sodium salt is a luminol-based chemiluminescence (CL) probe that is extensively used in vivo and in vitro to detect NADPH oxidase (Nox)-derived superoxide (O2•−) and identify Nox inhibitors.

Physicochemical Properties

Molecular Formula	C13H8CLN4NAO2
Molecular Weight	310.67
Exact Mass	310.023
CAS #	143556-24-5
PubChem CID	78243715
Appearance	White to yellow solid powder
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	1
Heavy Atom Count	21
Complexity	430
Defined Atom Stereocenter Count	0
InChi Key	IGEUYSJHQQCEFP-UHFFFAOYSA-M
InChi Code	InChI=1S/C13H9ClN4O2.Na/c14-11-8-7(12(19)17-18-13(8)20)9(15)10(16-11)6-4-2-1-3-5-6;/h1-5H,15H2,(H,17,19)(H,18,20);/q;+1/p-1
Chemical Name	sodium;8-amino-5-chloro-1-oxo-7-phenyl-2H-pyrido[3,4-d]pyridazin-4-olate
Synonyms	L-012 sodium; L012 sodium; L012 sodium salt
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.
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	L012 Sodium salt is a chemical analogue of Luminol that, when compared to MCLA and Lucigenin, other CL highlights, greatly enhances luminescence production.
ln Vivo	It is possible to track the growth and control of biosensors by utilizing the robust ROS/RNS-dependent luminescence signals that are mediated in vivo by the sodium salt L-012 [2], which is widely distributed in mice.
References	[1]. On the use of L-012, a luminol-based chemiluminescent probe, for detecting superoxide and identifying inhibitors of NADPH oxidase: a reevaluation. Free Radic Biol Med. 2013;65:1310-1314. [2]. In vivo imaging of reactive oxygen and nitrogen species in inflammation using the luminescent probe L-012. Free Radic Biol Med. 2009;47(6):760-766.

Solubility Data

Solubility (In Vitro)

DMSO : ~50 mg/mL (~160.94 mM) H2O : ~3.33 mg/mL (~10.72 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 1.25 mg/mL (4.02 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 (4.02 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	3.2188 mL	16.0942 mL	32.1885 mL
	5 mM	0.6438 mL	3.2188 mL	6.4377 mL
	10 mM	0.3219 mL	1.6094 mL	3.2188 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.