N-Ethylmaleimide (Synonyms: NEM)

Name: N-Ethylmaleimide
Brand: MedChemExpress
SKU: HY-D0843
Rating: 5.0 (37 reviews)

Cat. No.: HY-D0843 Purity: 99.59%: Data Sheet
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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.

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

CAS No. : 128-53-0

Size	Stock	Quantity

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Solid or liquid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO ready for reconstitution	In-stock	Estimated Time of Arrival: December 31
Solution
10 mM * 1 mL in DMSO	In-stock	Estimated Time of Arrival: December 31
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Based on 37 publication(s) in Google Scholar

Other Forms of N-Ethylmaleimide:

N-Ethylmaleimide-d₅ In-stock
N-Ethylmaleimide (Standard) Get quote

37 Publications Citing Use of MCE N-Ethylmaleimide

Cell Proliferation/Viability Assay

Flow Cytometry

In Vivo Efficacy Study

IHC

: N-Ethylmaleimide purchased from MedChemExpress. Usage Cited in: Theranostics. 2024 Jun 1;14(9):3470-3485. [Abstract]; MHCC97H SR and Hep3B SR cells were treated with sorafenib for 24 hours, with or without NEM (10 μM) or FSG67 (30 μM), and cell death was assessed by the CCK-8 assay.

: N-Ethylmaleimide purchased from MedChemExpress. Usage Cited in: Theranostics. 2024 Jun 1;14(9):3470-3485. [Abstract]; MHCC97H SR and Hep3B SR cells were treated with sorafenib for 24 hours, with or without NEM (10 μM) or FSG67 (30 μM), and apoptosis was assessed by flow cytometry.

: N-Ethylmaleimide purchased from MedChemExpress. Usage Cited in: Theranostics. 2024 Jun 1;14(9):3470-3485. [Abstract]; The sensitivity of MHCC97H SR cells to sorafenib, NEM (1 mg/kg, every 2 days), and FSG67 was evaluated in vivo. The figure shows a solid view of the tumor.

: N-Ethylmaleimide purchased from MedChemExpress. Usage Cited in: Theranostics. 2024 Jun 1;14(9):3470-3485. [Abstract]; The sensitivity of MHCC97H SR cells to sorafenib, NEM (1 mg/kg, every 2 days), and FSG67 was evaluated in vivo. The figure shows the tumor growth curve.

: N-Ethylmaleimide purchased from MedChemExpress. Usage Cited in: Theranostics. 2024 Jun 1;14(9):3470-3485. [Abstract]; The sensitivity of MHCC97H SR cells to sorafenib, NEM (1 mg/kg, every 2 days), and FSG67 was evaluated in vivo. The figure shows the results of p65 and Oil Red O staining immunohistochemical (IHC) analysis.

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USP1 USP2 USP4 USP7 USP8 USP10 USP21 USP30 UCHL1 USP11 USP15

Biological Activity
Purity & Documentation
References
Customer Review

Description

IC₅₀ & Target

IC50: 6.3 μM (prolyl endopeptidase)^[2]

Cellular Effect

Cell Line	Type	Value	Description	References
COS-7	EC₅₀	43.7 μM Compound: NEM	Binding affinity to rat Galpha-fused ORL1 receptor expressed in african green monkey COS7 cells after 90 mins Binding affinity to rat Galpha-fused ORL1 receptor expressed in african green monkey COS7 cells after 90 mins	[PMID: 22061823]
KB 3-1	IC₅₀	0.03 mM Compound: N-ethyl maleimide	Cytotoxicity against human KB-3-1 cells incubated for 72 hrs by MTT assay Cytotoxicity against human KB-3-1 cells incubated for 72 hrs by MTT assay	[PMID: 21657271]
KB-V1	IC₅₀	0.016 mM Compound: N-ethyl maleimide	Cytotoxicity against drug-resistant human KBV1 cells expressing P-gp incubated for 72 hrs by MTT assay Cytotoxicity against drug-resistant human KBV1 cells expressing P-gp incubated for 72 hrs by MTT assay	[PMID: 21657271]
MCF7	GI₅₀	9.9 μg/mL Compound: 45	Cytotoxicity against human MCF7 cells assessed as cell growth after 2 days by sulforhodamine B assay Cytotoxicity against human MCF7 cells assessed as cell growth after 2 days by sulforhodamine B assay	[PMID: 21481593]
NCI-H460	GI₅₀	6.7 μg/mL Compound: 45	Cytotoxicity against human H460 cells assessed as cell growth after 2 days by sulforhodamine B assay Cytotoxicity against human H460 cells assessed as cell growth after 2 days by sulforhodamine B assay	[PMID: 21481593]
SF-268	GI₅₀	6.5 μg/mL Compound: 45	Cytotoxicity against human SF268 cells assessed as cell growth after 2 days by sulforhodamine B assay Cytotoxicity against human SF268 cells assessed as cell growth after 2 days by sulforhodamine B assay	[PMID: 21481593]

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.

128-53-0

Appearance

<43°C Solid,>46°C Liquid

Color

White to light yellow

SMILES

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

Initial Source

Microorganisms

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 : 100 mg/mL (799.17 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

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

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
10 mM	0.7992 mL	3.9958 mL	7.9917 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)

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: ≥ 2.08 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.
Protocol 3

Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 2.08 mg/mL (16.62 mM); Clear solution

This protocol yields a clear solution of ≥ 2.08 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 (20.8 mg/mL) to 900 μL Corn oil, and mix evenly.

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)

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%

Select Batch:

Data Sheet (286 KB) SDS (642 KB)

COA (258 KB) LCMS (94 KB)

Handling Instructions (2659 KB)

References

[1]. Moriyama A, et al. Porcine muscle prolyl endopeptidase and its endogenous substrates. J Biochem. 1988 Jul;104(1):112-7. [Content Brief]

[2]. Yellaturu CR, et al. N-Ethylmaleimide inhibits platelet-derived growth factor BB-stimulated Akt phosphorylation via activation of protein phosphatase 2A. J Biol Chem. 2002 Oct 18;277(42):40148-55. [Content Brief]

[3]. Matsuda H, et al. Roles of capsaicin-sensitive sensory nerves, endogenous nitric oxide, sulfhydryls, and prostaglandins in gastroprotection by momordin Ic, an oleanolic acid oligoglycoside, on ethanol-induced gastric mucosal lesions in rats. Life Sci. 1999;65(2):PL27-32. [Content Brief]

Complete Stock Solution Preparation Table

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.

N-Ethylmaleimide Related Classifications

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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.

Keywords:

N-Ethylmaleimide128-53-0NEMCathepsinDeubiquitinaseApoptosisDUBsVSMCcysteine residuemodifysulfhydrylphosphate transportInhibitorinhibitorinhibit

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N-Ethylmaleimide (Synonyms: NEM)

Customer Review

Other Forms of N-Ethylmaleimide:

N-Ethylmaleimide Related Antibodies

37 Publications Citing Use of MCE N-Ethylmaleimide

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•Related Screening Libraries:

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•Related Proteins:

View All Cathepsin Isoform Specific Products:

View All Deubiquitinase Isoform Specific Products:

Biological Activity

Purity & Documentation

References

Customer Review

N-Ethylmaleimide Related Antibodies

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Complete Stock Solution Preparation Table

N-Ethylmaleimide Related Classifications

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