试剂                        
                
                                
                        
                            转染                        
                
                                
                        
                            荧光素酶                        
                
                                
                        
                            组合化学                        
                
                                
                        
                            化学                        
                
                                
                        
                            生物化学                        
                
                                
                        
                            基因                        
                
                                
                        
                            有机化学                        
                
                        
                    
            作者
            
                Tracy J Worzella,T. Allison,Pete Stecha,Mei Cong,Chad Zimprich,Kevin Kershner,Richard L. Somberg            
         
                    
            出处
            
                                    期刊:Cancer Research
                                                         [American Association for Cancer Research]
                                                        日期:2012-04-01
                                                        卷期号:72 (8_Supplement): 3883-3883
                                                 
         
        
    
            
            标识
            
                                    DOI:10.1158/1538-7445.am2012-3883
                                    
                                
                                 
         
        
                
            摘要
            
            Abstract As biological assays used for HTS and uHTS are miniaturized, it is critical to maintain assay sensitivity while minimizing reagent volumes and maximizing throughput. The GloSensor™ cAMP Assay provides an extremely sensitive and easy to use, real-time luminescent assay format for the interrogation of over expressed or endogenous GPCRs that signal via changes in the intracellular concentration of cAMP. Tipless, touchless transfers with the Echo® liquid handler eliminate the need for costly disposable tips and greatly simplify assay development efforts. Precise and accurate drop placement eliminates cross-contamination. In this work, we demonstrate optimization of the GloSensor cAMP Assay in low-volume format using a stably transfected HEK293 cell line model expressing the melanocortin 4 receptor (MC4R) and a cAMP-sensing variant of firefly luciferase. We show that the Echo liquid handler is able to titrate small molecule and peptide agonists and antagonists, as well as transfer the GloSensor cAMP Reagent, providing robust assay results that are achieved with significantly reduced volumes of cells and compound. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3883. doi:1538-7445.AM2012-3883
         
            
 
                 
                
                    
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