HADA HCl (HCC-Amino-D-alanine HCl) is a blue fluorescent FDAA (λem~450 nm). FDAAs efficiently bind to PGs of a variety of bacteria at the site of peptidoglycan (PG) synthesis, allowing specific and covalent detection of bacterial growth with minimal interference.

Physicochemical Properties

Molecular Formula	C13H13CLN2O6
Molecular Weight	328.705122709274
CAS #	2253733-10-5
Related CAS #	1417905-41-9;2253733-10-5 (HCl);
PubChem CID	137919867
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	5
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	4
Heavy Atom Count	22
Complexity	489
Defined Atom Stereocenter Count	1
SMILES	Cl.O1C(C(C(NC[C@H](C(=O)O)N)=O)=CC2C=CC(=CC1=2)O)=O
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	FDAA labeling for organisms that develop quickly (like E. Coli) only takes 30 seconds. HADA hydrochloride offers the most reliable and consistent PG labeling for the majority of bacterial species and typically does not require significant adjustment, despite being less photostable and darker than FDL or TDL [1].
References	[1]. Synthesis of fluorescent D-amino acids and their use for probing peptidoglycan synthesis and bacterial growth in situ. Nat Protoc. 2015 Jan;10(1):33-52.

Solubility Data

Solubility (In Vitro)

DMSO : 125 mg/mL (380.27 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 2.08 mg/mL (6.33 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 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: ≥ 2.08 mg/mL (6.33 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 20.8 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (6.33 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.0422 mL	15.2110 mL	30.4220 mL
	5 mM	0.6084 mL	3.0422 mL	6.0844 mL
	10 mM	0.3042 mL	1.5211 mL	3.0422 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.