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Volumn 20, Issue 22, 2010, Pages 6560-6564

Inhibition of Bfl-1 with N-aryl maleimides

Author keywords

Anti cancer agents; Bfl 1 inhibitors; High throughput screening; N Aryl maleimides

Indexed keywords

3,4 DICHLORO 1 (3,4 DICHLOROPHENYL) 1H PYROLE 2,5 DIONE; AMINE; ANTINEOPLASTIC AGENT; BCL2 RELATED PROTEIN A1; MALEIMIDE DERIVATIVE; UNCLASSIFIED DRUG;

EID: 77958043876     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2010.09.046     Document Type: Article
Times cited : (22)

References (21)
  • 17
    • 77958036242 scopus 로고    scopus 로고
    • note
    • Day, C. L.; Chen, L.; Richardson, S. J.; Harrison, P. J.; Huang, D. C. S.; Hings, M. G. J. Biol. Chem. 2005, 280. High-throughput FP assay: Chemical inhibitors were dissolved in 10% DMSO at 100 μM stock concentration. From this solution, 4 μL were incubated with 8 μL of Bfl-1 working solution (7.4 nM GST-Bfl-1 fusion protein in 25 mM Bis-Tris pH 7.0, 1 mM TCEP, 0.005% Tween 20) at room temperature away from direct sunlight in 384-well black plates (Greiner Bio-One, Monroe, NC). After 1 h, FITC-Bid BH3 peptide (8 μL of 5.6 nM in 25 mM Bis-Tris pH 7.0, 1 mM TCEP, 0.005% Tween 20) was added, and plates were incubated at room temperature away from direct sunlight. Fluorescence polarization was measured using an Analyst GT plate reader (Molecular Devices, Inc., Sunnyvale, CA) using fluorescein filters (excitation filter 485 nm, emission filter 530 nm, dichroic mirror # 505 nm). Data analysis was done using CBIS software (ChemInnovations, Inc., San Diego, CA). Dose-response confirmation: Chemical inhibitors were serially diluted in DMSO then in water so final concentration was in 10% DMSO. Ten concentrations of each compound were used. Each inhibitor (4 μL) was transferred to 384-well black plates (Greiner Bio-One, Monroe, NC) followed by GST-Bfl-1 solution described above (8 μL). Plates were incubated for 1 h at 4 °C. FITC-Bid BH3 peptide solution described above (8 μL) was added, and plates were incubated for 4 h at room temperature. Fluorescence polarization was measured at described above. Data analysis was performed using sigmoidal dose-response equation through non-linear regression.
    • (2005) J. Biol. Chem. , pp. 280
    • Day, C.L.1    Chen, L.2    Richardson, S.J.3    Harrison, P.J.4    Huang, D.C.S.5    Hings, M.G.6
  • 18
    • 77958037981 scopus 로고    scopus 로고
    • note
    • 50 data in the tables) can be found at: http://graphpad.com/help/prism5/prism5help.html? usingstatistical-analyses-step-by-s.htm.
  • 21
    • 77958070620 scopus 로고    scopus 로고
    • http://www.ncbi.nlm.nih.gov/pccompound.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.