L-Azidonorleucine HCl is an unnatural amino acid (AA) that is a substitute for Methionine. L-Azidonorleucine HCl may be utilized to label mammalian cell proteins and identify multiple methionyl-tRNA synthetase (MetRS) mutants. L-Azidonorleucine ( HCl) is a click chemical reagent. It has an azide (N3) moiety and could undergo CuAAc (copper-catalyzed azide-alkyne cycloaddition reaction) with compounds bearing an alkyne group. SPAAC (Strain-promoted alkyne-azide cycloaddition) may also happen with compounds bearing a BCN or DBCO group.

Physicochemical Properties

Molecular Formula	C6H13CLN4O2
Molecular Weight	208.65
Exact Mass	208.072
CAS #	1454334-76-9
PubChem CID	121235322
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	6
Heavy Atom Count	13
Complexity	188
Defined Atom Stereocenter Count	1
SMILES	C(CCN=[N+]=[N-])C[C@@H](C(=O)O)N.Cl
InChi Key	RCEAACZNVVRXSJ-JEDNCBNOSA-N
InChi Code	InChI=1S/C6H12N4O2.ClH/c7-5(6(11)12)3-1-2-4-9-10-8;/h5H,1-4,7H2,(H,11,12);1H/t5-;/m0./s1
Chemical Name	(2S)-2-amino-6-azidohexanoic acid;hydrochloride
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, 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	After being transfected with pMaRSC and treated for six hours with 1.5 mM of L-Azidonorleucine hydrochloride (AnI), CHO cells were permeabilized, fixed, and given alkyne-TAMRA treatment. Anl cells selectively incorporated cellular proteins, as evidenced by the observation of TAMRA fluorescence exclusively in mCherry-expressing cells [1].
ln Vivo	To label and identify neuronal populations in the brain, an average daily intake of L-Azidonorleucine (ANL) of 0.9 mg/day/g body weight over a period of 21 days is advised when ANL is delivered through drinking water. Other cell types can be labeled with lower doses or shorter labeling times. It is advised to experiment with various intraperitoneal injection doses of L-azidonorleucine within the 4-400 mM range. It is advised to use intraperitoneal injection; to label specific neuronal populations (such as the excitatory neurons in the hippocampus), a 400 mM dose (ip volume: 10 ml/kg) should be administered once daily for one week. When labeling different cell types or when longer labeling times are needed, lower doses—such as 4 mM—may be used [2].
References	[1]. Engineered Aminoacyl-tRNA Synthetase for Cell-Selective Analysis of Mammalian Protein Synthesis. J Am Chem Soc. 2016 Apr 6;138(13):4278-81. [2]. Cell-type-specific metabolic labeling, detection and identification of nascent proteomes in vivo. Nat Protoc. 2019 Feb;14(2):556-575.

Solubility Data

Solubility (In Vitro)	H2O: 125 mg/mL (599.09 mM)
Solubility (In Vivo)	Solubility in Formulation 1: 100 mg/mL (479.27 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	4.7927 mL	23.9636 mL	47.9272 mL
	5 mM	0.9585 mL	4.7927 mL	9.5854 mL
	10 mM	0.4793 mL	2.3964 mL	4.7927 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.