Furimazine-PBI-3939-DataSheet-生命科学试剂-MedChemExpress

1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEFurimazineCat. No.: HY-111497CAS No.: 1374040-24-0Synonyms: PBI 3939分式: CHNO分量: 381.43作靶点: Others作通路: Others储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 100 mg/mL (262.17 mM)* means solu

2、ble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制备储备液1 mM 2.6217 mL 13.1086 mL 26.2171 mL5 mM 0.5243 mL 2.6217 mL 5.2434 mL10 mM 0.2622 mL 1.3109 mL 2.6217 mL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，并请注意储备液的保存式和期限。BIOLOGICAL ACTIVITY物活性 Furimazine种咪唑并吡嗪酮类的发光底物。在哺乳动物细胞中， NanoLuc (Nluc) 与 Fur

3、imazine 联，产更亮的发光，带 Coelenterazine 的 Oluc-19 250 万倍。体外研究Furimazine is a Coelenterazine analogue.The apparent Km for purified NanoLuc (Nluc) using eitherFurimazine or Coelenterazine is 10 M, while the maximum luminescence (i.e., apparent Vmax) is 30-fold1/2 Master of Small Molecules 您边的抑制剂师www.MedChem

4、Ehigher for Furimazine than for native Coelenterazine. Measurement of luminescence intensity from cellsexpressing Nluc reveals that maximal signal is attained at about 10-20 M Furimazine 1.PROTOCOLKinase Assay 1 Random libraries are generated by error-prone PCR (average of 2-3 mutations per clone).

5、Library 1 (phase1; template=Oluc-N166R) is screened (4,400 variants) with Coelenterazine. Library 2 (phase 2;template=C1A4E) is screened (4,400 variants) with 11 novel Coelenterazine analogues: 3840, 3841, 3842,3857, 3880, 3881, 3886, 3887, 3889, 3897, and 3900. The 11 analogues represent substituti

6、ons at positions2, 6, and 8 and are considered to be representative of the entire set of 24 compounds; 2,200 variants arescreened with compounds 3896 and 3894. All hits (improved luminescence) are screened again with theremaining Coelenterazine analogues. Library 3 (phase 3;template=C1A4E+Q18L/K33N/

7、F54I/F68Y/L72Q/M75K/I90V) was screened in the context of a mouse Id-X-HaloTag (where X=library) using Coelenterazine and Furimazine. Library screens are performed on aFreedom robotic workstation as follows: induced bacterial cultures (in 96-well microtiter plates) are lysed witha buffer containing 3

8、00 mM HEPES pH 8, 200 mM thiourea, 0.3X Passive Lysis Buffer, 0.3 mg/mL lysozyme,and 0.002 units of RQ1 DNase. Assay reagent containing 1 mM CDTA, 150 mM KCl, 10 mM DTT, 0.5% (v/v)Tergitol, and 20 M substrate is then added to equal volumes of lysate. Samples are measured on a GENiosPro luminometer 1

9、.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Hall MP, et al. Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem Biol.2012 Nov 16;7(11):1848-57.McePdfHeightCaution: Product has not been fully validated for medical