Alkyne-phenol (Alk-Ph) is an ascorbate peroxidase 2 (APEX2) probe. Alkyne-phenol significantly improves APEX labeling efficiency of intact yeast cells and has greater cell wall permeability than the APEX2 substrate biotin-phenol (BP). Alkyne-phenol also helps identify APEX labeling sites and clarify the assignment of membrane topology to mitochondrial proteins. Alkyne-phenol is a reagent for click chemistry. It contains Alkyne groups and could undergo CuAAc (copper-catalyzed azide-alkyne cycloaddition reaction) with compounds bearing an Azide group.

Physicochemical Properties

Molecular Formula	C13H15NO2
Molecular Weight	217.263703584671
Exact Mass	217.11
CAS #	1694495-59-4
PubChem CID	108021005
Appearance	White to off-white solid powder
LogP	1.7
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	2
Rotatable Bond Count	5
Heavy Atom Count	16
Complexity	258
Defined Atom Stereocenter Count	0
SMILES	O=C(CCC#C)NCCC1C=CC(=CC=1)O
InChi Key	CQZJBJSXQYZLEX-UHFFFAOYSA-N
InChi Code	InChI=1S/C13H15NO2/c1-2-3-4-13(16)14-10-9-11-5-7-12(15)8-6-11/h1,5-8,15H,3-4,9-10H2,(H,14,16)
Chemical Name	N-[2-(4-hydroxyphenyl)ethyl]pent-4-ynamide
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

Each probe was added to yeast cells expressing Su9-APEX2 at a final concentration of 2.5 mM for 30 minutes of incubation. The labeling reaction was then started by adding 1 mM H2O2. The most effective APEX2 substrate for yeast proteome labeling is alkyne-phenol [1]. At probe concentrations greater than 1 mM, APEX2-mediated Alkyne-phenol labeling in yeast mitochondria is more successful [1]. Compared to the conventional APEX2 substrate biotin-phenol (BP), alkyne-phenol offers greater coverage and an exceptionally high specificity (94%) for proteomic investigation of the mitochondrial matrix of intact yeast cells [1].

References

[1]. A Clickable APEX Probe for Proximity-Dependent Proteomic Profiling in Yeast. Cell Chem Biol. 2020;27(7):858-865.e8.

Solubility Data

Solubility (In Vitro)

DMSO: 50 mg/mL (230.14 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (11.51 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 25.0 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: ≥ 2.5 mg/mL (11.51 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 25.0 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: ≥ 2.5 mg/mL (11.51 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 25.0 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	4.6028 mL	23.0139 mL	46.0278 mL
	5 mM	0.9206 mL	4.6028 mL	9.2056 mL
	10 mM	0.4603 mL	2.3014 mL	4.6028 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.