NEM (N-Ethylmaleimide)

Catalog No.S3692 Batch:S369202

Technical Data

Formula	C₆H₇NO₂
Molecular Weight	125.13	CAS No.	128-53-0
Solubility (25°C)*	In vitro	DMSO	25 mg/mL (199.79 mM)
		Water	25 mg/mL (199.79 mM)
		Ethanol	25 mg/mL (199.79 mM)
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

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	N-Ethylmaleimide inactivates endogenous deubiquitinating enzymes (DUBs) and specifically inhibits phosphate transport in mitochondria.^[1]
In vivo	N-Ethylmaleimide is an irreversible cysteine protease (sulfhydryl) inhibitor.

Protocol (from reference)

Cell Assay:^{^[1]}	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.
Animal Study:^{^[2]}	Animal Models Male Sprague-Dawley rats Dosages 10 mg/kg Administration s.c.

Cell Assay:^{^[1]}

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.

Animal Study:^{^[2]}

Animal Models
Male Sprague-Dawley rats
Dosages
10 mg/kg
Administration
s.c.

References

Selleck's NEM (N-Ethylmaleimide) has been cited by 11 publications

A combinatorial therapeutic approach to enhance FLT3-ITD AML treatment [ Cell Rep Med, 2023, 10.1016/j.xcrm.2023.101286]	PubMed: 37951217
Perturbation of Copper Homeostasis Sensitizes Cancer Cells to Elevated Temperature [ Int J Mol Sci, 2023, 25(1)423]	PubMed: 38203594
Targeting PELP1 oncogenic signaling in TNBC with the small molecule inhibitor SMIP34 [ Breast Cancer Res Treat, 2023, 200(1):151-162]	PubMed: 37199805
A First-in-Class Inhibitor of ER Coregulator PELP1 Targets ER+ Breast Cancer [ Cancer Res, 2022, 82-20:3830-3844]	PubMed: 35950923
Neddylation is essential for β-catenin degradation in Wnt signaling pathway [ Cell Rep, 2022, 38(12):110538]	PubMed: 35320710
The E3 Ligase RNF157 Inhibits Lens Epithelial Cell Apoptosis by Negatively Regulating p53 in Age-Related Cataracts [ Invest Ophthalmol Vis Sci, 2022, 63(4):11]	PubMed: 35435923
Histone chaperone CAF-1 promotes HIV-1 latency by leading the formation of phase-separated suppressive nuclear bodies [ EMBO J, 2021, 40(10):e106632]	PubMed: 33739466
Mitochondrial Drp1 recognizes and induces excessive mPTP opening after hypoxia through BAX-PiC and LRRK2-HK2 [ Cell Death Dis, 2021, 12(11):1050]	PubMed: 34741026
Ubiquitination of the DNA-damage checkpoint kinase CHK1 by TRAF4 is required for CHK1 activation. [ J Hematol Oncol, 2020, 1;13(1):40]	PubMed: 32357935
DIPK2A promotes STX17- and VAMP7-mediated autophagosome-lysosome fusion by binding to VAMP7B. [ Autophagy, 2019, 10.1080/15548627]	PubMed: 31251111

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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