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  3. Z-Gly-Pro-AMC

Z-Gly-Pro-AMC is a fluorogenic substrate. Z-Gly-Pro-AMC is hydrolyzed by prolyl endopeptidase to generate highly fluorescent 7-amido-4-methylcoumarin (HY-D0027). (λex=380 nm, λem=465 nm).

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

Z-Gly-Pro-AMC Chemical Structure

Z-Gly-Pro-AMC Chemical Structure

CAS No. : 68542-93-8

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Based on 1 publication(s) in Google Scholar

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  • Biological Activity

  • Purity & Documentation

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Description

Z-Gly-Pro-AMC is a fluorogenic substrate. Z-Gly-Pro-AMC is hydrolyzed by prolyl endopeptidase to generate highly fluorescent 7-amido-4-methylcoumarin (HY-D0027). (λex=380 nm, λem=465 nm)[1].

In Vitro

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Z-Gly-Pro-AMC assay[1]:
1. Make a 5 μL of plasma sample is pre-incubated with 10 μL of 250 nM FAP inhibitor, 10 μL of 250 nM PREP inhibitor or 10 μL of 0.0025% (v/v) DMSO for 15 min at 37 ℃.
2. Add 35 μL pre-heated Z-Gly-Pro-AMC (380 μM diluted in buffer) to obtain a final concentration of 266 μM.
3. Fluorescence is measured kinetically for 30 min at 37 ℃.

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

Molecular Weight

463.48

Formula

C25H25N3O6

CAS No.
Appearance

Solid

Color

White to off-white

Emission (Em)

460

Excitation (Ex)

380

SMILES

CC(C1=CC=C(NC([C@H]2N(CCC2)C(CNC(OCC3=CC=CC=C3)=O)=O)=O)C=C1O4)=CC4=O

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

-20°C, protect from light, stored under nitrogen

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

Solvent & Solubility
In Vitro: 

DMSO : 125 mg/mL (269.70 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 2.1576 mL 10.7880 mL 21.5759 mL
5 mM 0.4315 mL 2.1576 mL 4.3152 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, stored under nitrogen). 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)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

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

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

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: ≥ 2.08 mg/mL (4.49 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 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: ≥ 2.08 mg/mL (4.49 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 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.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

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, stored under nitrogen)

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: 99.80%

References

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, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 2.1576 mL 10.7880 mL 21.5759 mL 53.9398 mL
5 mM 0.4315 mL 2.1576 mL 4.3152 mL 10.7880 mL
10 mM 0.2158 mL 1.0788 mL 2.1576 mL 5.3940 mL
15 mM 0.1438 mL 0.7192 mL 1.4384 mL 3.5960 mL
20 mM 0.1079 mL 0.5394 mL 1.0788 mL 2.6970 mL
25 mM 0.0863 mL 0.4315 mL 0.8630 mL 2.1576 mL
30 mM 0.0719 mL 0.3596 mL 0.7192 mL 1.7980 mL
40 mM 0.0539 mL 0.2697 mL 0.5394 mL 1.3485 mL
50 mM 0.0432 mL 0.2158 mL 0.4315 mL 1.0788 mL
60 mM 0.0360 mL 0.1798 mL 0.3596 mL 0.8990 mL
80 mM 0.0270 mL 0.1348 mL 0.2697 mL 0.6742 mL
100 mM 0.0216 mL 0.1079 mL 0.2158 mL 0.5394 mL
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Z-Gly-Pro-AMC Related Classifications

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  • 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.

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Z-Gly-Pro-AMC
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