MCA (Synonyms: 7-Methoxycoumarin-4-acetic acid)

Cat. No.: HY-W027544 Purity: 98.87%: FAQs
Data Sheet
SDS

COA
Handling Instructions Technical Support

MCA (7-Methoxycoumarin-4-acetic acid) is a Coumarin (HY-N0709) derivative. MCA quantitates platelet-activating factor (PAF) by high-performance liquid chromatography with fluorescent detection. MCA can modify FRET peptide substrates for analyzing protease activities.

For research use only. We do not sell to patients.

MCA Chemical Structure

CAS No. : 62935-72-2

Size	Stock	Quantity
100 mg	In-stock	Estimated Time of Arrival: December 31
250 mg	In-stock	Estimated Time of Arrival: December 31
500 mg	In-stock	Estimated Time of Arrival: December 31
1 g	Get quote
5 g	Get quote

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Other Forms of MCA:

MCA-d₃ Get quote
Dansyl propargylamine-d₆ Get quote

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References
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Description

In Vitro

1. FRET preparation^[1]
1) Synthesize FRET substrates on TCP resin using Fmoc/tBu chemistry by fully automated solid-phase peptide synthesis.
2) Use a 7-fold molar excess of single Fmoc-L-amino acids for the coupling of standard D- or L-amino acids.
3) Perform Fmoc deprotections twice, each for 10 min, with 30% piperidine in dimethylformamide.
4) Couple Nα-Fmoc-N-ε-2,4-DNP-L-lysine [Fmoc-Lys(Dnp)] in 2-fold excess within 2 h with 1-H-benzotriazolium-1-[bis(dimethylamino) methylene]-5-chloro-tetrafluoro-borate(1-)3-oxide/Oxima Pure.
5) Modify the peptide resin with 7-methoxycoumarin-4-acetic acid (MCA). Carry out the coupling with five equivalents of MCA in dimethylformamide with diisopropylcarbodiimide/Oxima Pure within 3 h and monitor by Kaiser assay.
6) Cleave the FRET substrates off the resin within 4 h by treating the resins with trifluoroacetic acid/phenol/ethanedithiol/thioanisole/water (95:2:1:1:1), and then precipitate and wash them three times with diethylether.
7) Lyophilize the peptides in tert-butyl alcohol/water (4:1).
8) Characterize the FRET substrates analytically by reversed-phase HPLC-electrospray mass spectrometry to ensure the compound purity is greater than 85% (UV 214 nm).
9) Prepare the working solution with the prepared FRET substrates according to experimental requirements. For example, in some experiments, the final concentration of the FRET substrate is prepared to be 0.5 mM (when incubated with proteases).
2. Enzyme activity analysis
1) Produce recombinant catalytically active NSP4 (such as S-tag NSP4).
2) Add NSP4, NE, PR3, or CG at a final concentration of 6 mM to a commercial peptide library containing 0.5 mM FRET substrates in total.
3) Generate fluorescent signals by excitation at 320 nm and record them at 405 nm emission wavelength.
4) Only use the initial linear portion of the progress curve to calculate the enzymatic activity, expressed as relative fluorescence units per minute (increase of relative fluorescence units per minute). Set the value of the substrate that is cleaved best by the tested proteases to 100%, and present the average values with standard deviations as a percentage of this maximum value.
5) Analyze the cleaved fragments by liquid chromatography-tandem mass spectrometry after incubating for 5 and 30 min.

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

Clinical Trial

Molecular Weight

234.21

Formula

C₁₂H₁₀O₅

CAS No.

62935-72-2

Appearance

Solid

Color

White to off-white

Emission (Em)

450

Excitation (Ex)

345

SMILES

O=C1OC(C=C(OC)C=C2)=C2C(CC(O)=O)=C1

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Solvent & Solubility

In Vitro:

DMSO : 25 mg/mL (106.74 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	4.2697 mL	21.3484 mL	42.6967 mL
5 mM	0.8539 mL	4.2697 mL	8.5393 mL
10 mM	0.4270 mL	2.1348 mL	4.2697 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 (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Molarity Calculator
Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

Concentration _(start)

Volume _(start)

Concentration _(final)

Volume _(final)

In Vivo:

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 (10.67 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% Corn Oil
Solubility: ≥ 2.5 mg/mL (10.67 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

DMSO +

Tween-80 +

Saline

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Calculation results:

Working solution concentration: mg/mL

Method for preparing stock solution: mg drug dissolved in μL DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).

Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.

If the continuous dosing period exceeds half a month, please choose this protocol carefully.

Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.

Purity & Documentation

Purity: 98.87%

Select Batch:

Data Sheet (276 KB) SDS (393 KB)

COA (250 KB) HNMR (112 KB) RP-HPLC (230 KB)

Handling Instructions (2659 KB)

References

[1]. Perera NC, et, al. NSP4 is stored in azurophil granules and released by activated neutrophils as active endoprotease with restricted specificity. J Immunol. 2013 Sep 1;191(5):2700-7. [Content Brief]

[2]. Choi YH,et, al. Effect of functional groups on the solubilities of coumarin derivatives in supercritical carbon dioxide. Chromatographia. 1998 Jan;47(93-7).

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

DMSO	1 mM	4.2697 mL	21.3484 mL	42.6967 mL	106.7418 mL
	5 mM	0.8539 mL	4.2697 mL	8.5393 mL	21.3484 mL
	10 mM	0.4270 mL	2.1348 mL	4.2697 mL	10.6742 mL
	15 mM	0.2846 mL	1.4232 mL	2.8464 mL	7.1161 mL
	20 mM	0.2135 mL	1.0674 mL	2.1348 mL	5.3371 mL
	25 mM	0.1708 mL	0.8539 mL	1.7079 mL	4.2697 mL
	30 mM	0.1423 mL	0.7116 mL	1.4232 mL	3.5581 mL
	40 mM	0.1067 mL	0.5337 mL	1.0674 mL	2.6685 mL
	50 mM	0.0854 mL	0.4270 mL	0.8539 mL	2.1348 mL
	60 mM	0.0712 mL	0.3558 mL	0.7116 mL	1.7790 mL
	80 mM	0.0534 mL	0.2669 mL	0.5337 mL	1.3343 mL
	100 mM	0.0427 mL	0.2135 mL	0.4270 mL	1.0674 mL

MCA Related Classifications

Others

Help & FAQs

Do most proteins show cross-species activity?
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

MCA62935-72-27-Methoxycoumarin-4-acetic acidFluorescent Dyecoumarinplatelet-activating factorhigh-performance liquid chromatographyFRET peptideproteaseInhibitorinhibitorinhibit

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MCA (Synonyms: 7-Methoxycoumarin-4-acetic acid)

Other Forms of MCA:

MCA Related Antibodies

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•Related Small Molecules:

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