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> Alkyne-phenol [1694495-59-4]

Alkyne-phenol [1694495-59-4]

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Cat# HY-131442-50mg

Size : 50mg

Brand : MedChemExpress

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Description

Alkyne-phenol (Alk-Ph) is a clickable ascorbate peroxidase 2 (APEX2) probe. Alkyne-phenol substantially improves APEX-labeling efficiency in intact yeast cells, as it is more cell wall-permeant than APEX2 substrate biotin-phenol (BP). Alkyne-phenol also facilitates the identification of APEX-labeling sites, allowing the unambiguous assignment of membrane topology of mitochondrial proteins[1]. Alkyne-phenol is a click chemistry reagent, it contains an Alkyne group and can undergo copper-catalyzed azide-alkyne cycloaddition (CuAAc) with molecules containing Azide groups.

In Vitro

Yeast cells expressing Su9-APEX2 are incubated with each probe at 2.5 mM final concentration for half an hour, and the labeling reaction is initiated by the addition of 1 mM H2O2. Alkyne-phenol is the most reactive APEX2 substrate for labeling the yeast proteome[1].
APEX2-mediated Alkyne-phenol labeling in the yeast mitochondria is more effective at probe concentrations above 1 mM[1].
Alkyne-phenol enables proteomic profiling in the mitochondrial matrix of intact yeast cells with exceptionally high specificity (94%), and offers higher coverage than the traditional APEX2 substrate biotin-phenol (BP)[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Alkyne-phenol Related Antibodies

Solvent & Solubility
In Vitro: 

DMSO : 50 mg/mL (230.14 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 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
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.5 mg/mL (11.51 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.5 mg/mL (11.51 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Purity & Documentation
References

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