Home Proteases Cysteine Protease inhibitor - DUB inhibitor NEM (N-Ethylmaleimide) For research use only.

NEM (N-Ethylmaleimide)

NEM (N-Ethylmaleimide) is an organic compound that is derived from maleic acid. It is an irreversible inhibitor of all cysteine peptidases, with alkylation occurring at the active site thiol group. N-Ethylmaleimide (NEM) inactivates endogenous deubiquitinating enzymes (DUBs). N-Ethylmaleimide (NEM) specifically inhibits phosphate transport in mitochondria.

NEM (N-Ethylmaleimide) Chemical Structure

NEM (N-Ethylmaleimide) Chemical Structure

CAS No. 128-53-0

Purity & Quality Control

NEM (N-Ethylmaleimide) Related Products

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
KBV1 Cytotoxicity assay 72 hrs Cytotoxicity against drug-resistant human KBV1 cells expressing P-gp incubated for 72 hrs by MTT assay, IC50=16μM 21657271
KB-3-1 Cytotoxicity assay 72 hrs Cytotoxicity against human KB-3-1 cells incubated for 72 hrs by MTT assay, IC50=30μM 21657271
COS7 Function assay 90 mins Binding affinity to rat Galpha-fused ORL1 receptor expressed in african green monkey COS7 cells after 90 mins, EC50=43.7μM 22061823
BL21 (DE3) Function assay 30 mins Irreversible inhibition of human recombinant IDE expressed in Escherichia coli BL21 (DE3) cells using ATTO 655- Cys-Lys-Leu-Val-Phe-Phe-Ala-Glu-Asp-Trp as substrate preincubated for 30 mins followed by 100 fold compound dilution by spectrophotometric anal 31276900
KU812 Function assay 20 mins Inhibition of SENP1 in human KU812 cells preincubated for 20 mins followed by addition of HA-SUMO1-VS as substrate measured after 2 hrs by immunoblot analysis 27491710
Click to View More Cell Line Experimental Data

Biological Activity

Description NEM (N-Ethylmaleimide) is an organic compound that is derived from maleic acid. It is an irreversible inhibitor of all cysteine peptidases, with alkylation occurring at the active site thiol group. N-Ethylmaleimide (NEM) inactivates endogenous deubiquitinating enzymes (DUBs). N-Ethylmaleimide (NEM) specifically inhibits phosphate transport in mitochondria.
In vitro
In vitro

N-Ethylmaleimide inactivates endogenous deubiquitinating enzymes (DUBs) and specifically inhibits phosphate transport in mitochondria.[1]
Cell Research Cell lines HEK293T cells
Concentrations 10 mM
Incubation Time 14 h
Method

For endogenous protein immunoprecipitation, cell lysates are prepared in Pierce IP Lysis Buffer containing protease inhibitors, phosphatase inhibitor and deubiquitinating inhibitor N-Ethylmaleimide (NEM, 10mM). Lysate supernatant is firstly incubated with primary antibody at 4 ℃ for 14 h, and then mixed with Dynabeads Protein G magnetic beads at 4 ℃ for 4 h on a rotator. The magnetic bead-Ab complex is separated by Magnetic Separation Rack and the beads are washed three times with PBST buffer. For SDS-PAGE analysis, the complex-bound beads are resuspended in 1x protein electrophoresis loading buffer and boiled at 100 ℃ for 8 min before the supernatant is collected. For exogenous protein immunoprecipitation, cells are transfected for 48 h before preparation of cell lysates. To prepare negative control, antibody against mouse IgG is used in parallel.
In Vivo
In vivo

N-Ethylmaleimide is an irreversible cysteine protease (sulfhydryl) inhibitor.
Animal Research Animal Models Male Sprague-Dawley rats
Dosages 10 mg/kg
Administration s.c.
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT00750477 Completed
Osteoarthritis
ESM Technologies LLC
 December 2004 Not Applicable
NCT00750230 Completed
Osteoarthritis|Fibromyalgia
ESM Technologies LLC
 October 2003 Not Applicable

Chemical Information & Solubility

Molecular Weight 125.13 Formula

C6H7NO2
CAS No. 128-53-0 SDF Download NEM (N-Ethylmaleimide) SDF
Smiles CCN1C(=O)C=CC1=O
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 25 mg/mL ( (199.79 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : 25 mg/mL

Ethanol : 25 mg/mL


Molecular Weight Calculator

In vivo
Batch:

Add solvents to the product individually and in order.


In vivo Formulation Calculator

Preparing Stock Solutions

Molarity Calculator

Mass Concentration Volume Molecular Weight

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
If you have any other enquiries, please leave a message.

* Indicates a Required Field

Please enter your name.
Please enter your email. Please enter a valid email address.
Please write something to us.
Tags: buy NEM (N-Ethylmaleimide) | NEM (N-Ethylmaleimide) supplier | purchase NEM (N-Ethylmaleimide) | NEM (N-Ethylmaleimide) cost | NEM (N-Ethylmaleimide) manufacturer | order NEM (N-Ethylmaleimide) | NEM (N-Ethylmaleimide) distributor