N-Ethylmaleimide
(Synonyms: N-乙基马来酰亚胺; NEM) 目录号 : GC34682
A modifier of protein sulfhydryl groups
Cas No.:128-53-0
Sample solution is provided at 25 µL, 10mM.
;)
,-1-7$$$37316672.jpg');)
;)
;)
$$$36806353.jpg');)
;33(7)%EF%BC%9A546-561$$$37156877.jpg');)
;)
;)
;)
%201124-1138.$$$35908550.jpg');)
%20766-780.$$$37100057.jpg');)
%20418-432.$$$36893736.jpg');)
%EF%BC%9A-240-255.$$$35108512.jpg');)
533-545$$$36543525.jpg');)
%201-13.$$$36600053.jpg');)
;)
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
N-Ethylmaleimide is a modifier of sulfhydryl groups in proteins.1,2 It has commonly been used to inactivate cysteine-containing enzymes in vitro. N-Ethylmaleimide inhibits prolyl endopeptidase with an IC50 value of 6.3 ?M.3 It alkylates cysteine 519 (Cys519) in voltage-gated potassium channel 7.4 (Kv7.4) and activates Kv7.2 , Kv7.4, and Kv7.5, but not Kv7.3, in CHO cells expressing the human channels when used at a concentration of 50 ?M.4
1.Kawakita, M., and Yamashita, T.Reactive sulfhydryl groups of sarcoplasmic reticulum ATPase. III. Identification of cysteine residues whose modification with N-ethylmaleimide leads to loss of the Ca2+-transporting activityJ. Biochem.102(1)103-109(1987) 2.Fatania, H.R., al-Nassar, K.E., and Thomas, N.Chemical modification of rat liver cytosolic NADP+-linked isocitrate dehydrogenase by N-ethylmaleimide. Evidence for essential sulphydryl groupsFEBS Lett.322(3)245-248(1993) 3.Moriyama, A., Nakanishi, M., and Sasaki, M.Porcine muscle prolyl endopeptidase and its endogenous substratesJ. Biochem.104(1)112-117(1988) 4.Li, Y., Gamper, N., and Shapiro, M.S.Single-channel analysis of KCNQ K+ channels reveals the mechanism of augmentation by a cysteine-modifying reagentJ. Neurosci.24(22)5079-5090(2004)
| Cas No. | 128-53-0 | SDF | |
| 别名 | N-乙基马来酰亚胺; NEM | ||
| Canonical SMILES | O=C(C=C1)N(CC)C1=O | ||
| 分子式 | C6H7NO2 | 分子量 | 125.13 |
| 溶解度 | DMSO : 50 mg/mL (399.58 mM) Water : 50 mg/mL (399.58 mM), Ethanol : 12.5 mg/mL (99.90 mM) | 储存条件 | Store at 4°C, sealed storage, away from moisture and light |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
||
| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
 |
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 |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Nociceptive behavior induced by the endogenous opioid peptides dynorphins in uninjured mice: evidence with intrathecal N-Ethylmaleimide inhibiting dynorphin degradation
Int Rev Neurobiol 2009;85:191-205.PMID:19607971DOI:10.1016/S0074-7742(09)85015-0.
Dynorphins, the endogenous opioid peptides derived from prodynorphin may participate not only in the inhibition, but also in facilitation of spinal nociceptive transmission. However, the mechanism of pronociceptive dynorphin actions, and the comparative potential of prodynorphin processing products to induce these actions were not fully elucidated. In our studies, we examined pronociceptive effects of prodynorphin fragments dynorphins A and B and big dynorphin consisting of dynorphins A and B, and focused on the mechanisms underlying these effects. Our principal finding was that big dynorphin was the most potent pronociceptive dynorphin; when administered intrathecally into mice at extremely low doses (1-10fmol), big dynorphin produced nociceptive behavior through the activation of the NMDA receptor ion-channel complex by acting on the polyamine recognition site. We next examined whether the endogenous dynorphins participate in the spinal nociceptive transmission using N-Ethylmaleimide (NEM) that blocks dynorphin degradation by inhibiting cysteine proteases. Similar to big dynorphin and dynorphin A, NEM produced nociceptive behavior mediated through inhibition of the degradation of endogenous dynorphins, presumably big dynorphin that in turn activates the NMDA receptor ion-channel complex by acting on the polyamine recognition site. Our findings support the notion that endogenous dynorphins are critical neurochemical mediators of spinal nociceptive transmission in uninjured animals. This chapter will review above-described phenomena and their mechanism.
Incorporation of N-Ethylmaleimide into the membrane-bound ADP/ATP translocator. Isolation of the protein labeled with N-[3H]ethylmaleimide
Eur J Biochem 1982 Feb;122(1):133-9.PMID:6277630DOI:10.1111/j.1432-1033.1982.tb05858.x.
The incorporation of N-Ethylmaleimide into the 30,000-Mr component of beef-heart mitochondria has been studied as a function of various ligands to the ADP/ATP carrier and the isolation of the N-ethylmaleimide-labeled protein is reported. 1. The incorporation of N-Ethylmaleimide into the 30,000-Mr component is specifically stimulated by ADP and ATP. Thus by differential incorporation of N-Ethylmaleimide, the 30,000-Mr component is preferentially labeled. 2. Addition of carboxyatractylate inhibits, whereas bongkrekate tolerates, the incorporation of N-Ethylmaleimide. 3. After solubilization by Triton the purification of N-ethylmaleimide-labeled protein is facilitated in the presence of bongkrekate but not of carboxyatractylate, in agreement with the postulated existence of only a bongkrekate-N-ethylmaleimide-protein complex. The labeled protein was purified to homogeneity on hydroxyapatite in Triton and subsequently, after denaturation in dodecylsulfate, on Sepharose 6B. 4. The identify of the isolated labeled protein with the formerly isolated bongkrekate-protein or carboxyatractylate-protein complexes is confirmed by the isoelectric point and amino acid composition. 5. Two moles of N-Ethylmaleimide must be incorporated into the 30,000-Mr component in order to inhibit fully the binding of one mole carboxyatractylate. This corresponds to one -SH group per unit.
Investigation of an artifact during non-reduced capillary electrophoresis with sodium dodecyl sulfate analysis utilizing N-Ethylmaleimide as an alkylation reagent
Anal Biochem 2022 Oct 15;655:114833.PMID:35961398DOI:10.1016/j.ab.2022.114833.
This manuscript describes the formation of an artifact shoulder peak with a slightly larger retention time than the main peak under the standard non-reduced capillary electrophoresis with sodium dodecyl sulfate (nrCE-SDS) analysis of a therapeutic recombinant protein X, and clarifies the formation mechanism of the artifact caused by N-Ethylmaleimide (NEM) during the sample preparation procedure. A design of experiment (DoE) approach was used to investigate the impact of the factors on the formation of the impurity. Additionally, orthogonal analytical experiments were performed to study the root cause of this phenomenon. The results consistently suggested that the Michael addition reaction between NEM and lysine residues in protein X, and decreased electrophoretic mobility due to increased molecular weight, was the root cause for the artifact, which could be partially inhibited by modifications of incubation conditions. Thus, before performing the nrCE-SDS method, the effects of alkylation reagents and sample preparation procedure on analytical results need to be considered seriously.
N-Ethylmaleimide Dissociates α7 ACh Receptor from a Complex with NSF and Promotes Its Delivery to the Presynaptic Membrane
Neurochem Res 2016 Aug;41(8):2043-8.PMID:27105867DOI:10.1007/s11064-016-1915-z.
N-Ethylmaleimide (NEM)-sensitive factor (NSF) associates with soluble NSF attachment protein (SNAP), that binds to SNAP receptors (SNAREs) including syntaxin, SNAP25, and synaptobrevin. The complex of NSF/SNAP/SNAREs plays a critical role in the regulation of vesicular traffic. The present study investigated NEM-regulated α7 ACh receptor translocation. NSF associated with β-SNAP and the SNAREs syntaxin 1 and synaptobrevin 2 in the rat hippocampus. NSF also associated with the α7 ACh receptor subunit, the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor subunits GluA1 and GluA2, and the γ-aminobutyric acid A (GABAA) receptor γ2 subunit. NEM, an inhibitor of NSF, significantly dissociated the α7 ACh receptor subunit from a complex with NSF and increased cell surface localization of the receptor subunit, but such effect was not obtained with the GluA1, GluA2 or γ2 subunits. NEM, alternatively, dissociated synaptobrevin 2 from an assembly of NSF/β-SNAP/syntaxin 1/synaptobrevin 2. NEM significantly increased the rate of nicotine-triggered AMPA receptor-mediated miniature excitatory postsynaptic currents, without affecting the amplitude, in rat hippocampal slices. The results of the present study indicate that NEM releases the α7 ACh receptor subunit and synaptobrevin 2 from an assembly of α7 ACh receptor subunit/NSF/β-SNAP/syntaxin 1/synaptobrevin 2, thereby promoting delivery of the α7 ACh receptor subunit to presynaptic membrane.
N-Ethylmaleimide INHIBITION OF HORSESHOE CRAB HEMOCYTE AGGLUTINATION
Science 1964 May 29;144(3622):1147-8.PMID:14148439DOI:10.1126/science.144.3622.1147.
The sulfhydryl inhibitor N-Ethylmaleimide (NEM) inhibits the agglutination of horseshoe crab hemocytes. The inhibitory action is neutralized by adding cysteine to the NEM before it is mixed with the hemolymph. This new method for arresting horseshoe crab hemocyte agglutination suggests the participation of sulfhydryl groups in the process.