1. Metabolic Enzyme/Protease Cell Cycle/DNA Damage Apoptosis
  2. Cathepsin Deubiquitinase Apoptosis
  3. N-Ethylmaleimide

N-Ethylmaleimide  (Synonyms: NEM)

Cat. No.: HY-D0843 Purity: 99.89%
COA Handling Instructions

N-Ethylmaleimide (NEM) derives from maleic acid, it can alkylates free sulfhydryl. N-Ethylmaleimide is an irreversible cysteine protease inhibitor. N-ethylmaleimide specific inhibits phosphate transport in mitochondria. N-Ethylmaleimide inhibits prolyl endopeptidase with an IC50 value of 6.3 μM. N-Ethylmaleimide can be used to modify cysteine residues in proteins and peptides.

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N-Ethylmaleimide Chemical Structure

N-Ethylmaleimide Chemical Structure

CAS No. : 128-53-0

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10 mM * 1 mL in DMSO
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Customer Review

Based on 10 publication(s) in Google Scholar

Other Forms of N-Ethylmaleimide:

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  • Biological Activity

  • Purity & Documentation

  • References

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Description

N-Ethylmaleimide (NEM) derives from maleic acid, it can alkylates free sulfhydryl. N-Ethylmaleimide is an irreversible cysteine protease inhibitor. N-ethylmaleimide specific inhibits phosphate transport in mitochondria. N-Ethylmaleimide inhibits prolyl endopeptidase with an IC50 value of 6.3 μM. N-Ethylmaleimide can be used to modify cysteine residues in proteins and peptides[1][2][3].

IC50 & Target

IC50: 6.3 μM (prolyl endopeptidase)[2]

In Vitro

N-Ethylmaleimide (20 μM;30 min) inhibits Akt Ser-473, Akt Thr-308 , p70S6K, ribosomal protein S6, 4E-BP1, eIF4E, BAD and FKHR-L1 phosphorylation[2].
N-Ethylmaleimide (20 μM;30 min) affects conversion of pro-caspase-3 in vascular smooth muscle cells[2].
N-Ethylmaleimide (20 μM;6 h) promotes vascular smooth muscle cells apoptosis[2].
N-Ethylmaleimide (20 μM;30 min) affects PP2A activity and ROS production in vascular smooth muscle cells[2].

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

Western Blot Analysis[2]

Cell Line: Vascular smooth muscle cells
Concentration: 20 μM
Incubation Time: 2 hours
Result: Effectively inhibited platelet-derived growth factor-BB (PDGF-BB)-stimulated Akt Ser-473 , Akt Thr-308, p70S6K, ribosomal protein S6, 4E-BP1, BAD and FKHR-L1 phosphorylation with a concentration of 20 μM.

Western Blot Analysis[2]

Cell Line: Vascular smooth muscle cells
Concentration: 20 μM
Incubation Time: 2 hours
Result: Increased of 1.8-fold in the conversion of pro-caspase-3 into active form, and showed better effect with 20 ng/ml PDGF-BB adding.

Apoptosis Analysis[2]

Cell Line: Vascular smooth muscle cells
Concentration: 20 μM
Incubation Time: 6 hours
Result: Induced vascular smooth muscle cells apoptosis by 3-fold, and exhibited 5-fold apoptosis with 20 ng/ml PDGF-BB adding.

Cell Viability Assay[2]

Cell Line: Vascular smooth muscle cells
Concentration: 20 μM
Incubation Time: 30 min
Result: Increased PP2A activity 1.7-flod and increased ROS production 2-fold in vascular smooth muscle cells.
In Vivo

N-Ethylmaleimide (10 mg/kg; i.h.) promotes the prevalence situation of mice with acute gastric ulcers[3].

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

Animal Model: Male Wistar rats with acute gastric ulcers induced by absolute ethanol injection[3]
Dosage: 10 mg/kg
Administration: Subcutaneous injection; 10 mg/kg once
Result: Increased the lesion area of acute gastric ulcers and attenuated the gastroprotective effect of PAG in rats.
Clinical Trial
Molecular Weight

125.13

Formula

C6H7NO2

CAS No.
Appearance

<43°C Solid,>46°C Liquid

Color

White to light yellow

SMILES

O=C(C=C1)N(CC)C1=O

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: 

H2O : 50 mg/mL (399.58 mM; Need ultrasonic)

DMSO : 50 mg/mL (399.58 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Ethanol : 12.5 mg/mL (99.90 mM; Need ultrasonic)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 7.9917 mL 39.9584 mL 79.9169 mL
5 mM 1.5983 mL 7.9917 mL 15.9834 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.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

  • Molarity Calculator

  • Dilution Calculator

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

Mass
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Concentration
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Volume
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Molecular Weight *

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

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

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Volume (start)

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C2

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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 (16.62 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 (16.62 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.

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
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:  PBS

    Solubility: 100 mg/mL (799.17 mM); Clear solution; Need ultrasonic

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.
Calculation results:
Working solution concentration: mg/mL
This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.
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).
Purity & Documentation

Purity: 99.89%

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). 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
Ethanol / H2O / DMSO 1 mM 7.9917 mL 39.9584 mL 79.9169 mL 199.7922 mL
5 mM 1.5983 mL 7.9917 mL 15.9834 mL 39.9584 mL
10 mM 0.7992 mL 3.9958 mL 7.9917 mL 19.9792 mL
15 mM 0.5328 mL 2.6639 mL 5.3278 mL 13.3195 mL
20 mM 0.3996 mL 1.9979 mL 3.9958 mL 9.9896 mL
25 mM 0.3197 mL 1.5983 mL 3.1967 mL 7.9917 mL
30 mM 0.2664 mL 1.3319 mL 2.6639 mL 6.6597 mL
40 mM 0.1998 mL 0.9990 mL 1.9979 mL 4.9948 mL
50 mM 0.1598 mL 0.7992 mL 1.5983 mL 3.9958 mL
60 mM 0.1332 mL 0.6660 mL 1.3319 mL 3.3299 mL
80 mM 0.0999 mL 0.4995 mL 0.9990 mL 2.4974 mL
H2O / DMSO 100 mM 0.0799 mL 0.3996 mL 0.7992 mL 1.9979 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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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.

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N-Ethylmaleimide
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