G蛋白偶聯(lián)受體（GPCR）是藥品發(fā)現(xiàn)計劃中大的靶向受體之一。鈣通量（通過Gq途徑耦合）分析是篩選GPCR靶點的方法。然而，超過60%的已知GPCRs信號通過腺苷酸環(huán)化酶活性偶聯(lián)到cAMP?，F(xiàn)有的cAMP檢測方法不僅需要細胞裂解，而且缺乏時間和空間分辨率。Screen Quest 活細胞cAMP檢測以高通量的形式提供細胞內(nèi)cAMP變化的實時監(jiān)測，無需細胞裂解步驟。該方法通過含有外源性環(huán)核苷酸門控通道（CNGC）或混雜G蛋白Gα16的細胞系進行。該通道被細胞內(nèi)cAMP水平升高，導(dǎo)致離子通量和細胞膜去極化，可通過熒光鈣（如Calbryte 520 AM、Cal-520 AM、Fluo-8 AM或Fluo-4 AM和相應(yīng)的免洗鈣試劑盒）或熒光膜電位染料檢測到。Gα16與特異性非Gq偶聯(lián)受體的共表達將在受體刺激下產(chǎn)生細胞內(nèi)鈣信號。Screen Quest 活細胞cAMP檢測提供細胞系和試劑，用于使用FLIPR、FDSS或其他等效的熒光酶標儀檢測細胞內(nèi)cAMP的變化。它已成功地用于測定Gs和Gi偶聯(lián)GPCR活性。百螢生物是AAT Bioquest的中國代理商，為您提供的Screen Quest 活細胞cAMP檢測。 

參考文獻

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