Pgp-Glo™试剂盒检测化合物对膜制备物中重组人类Pgp 活性的影响。该试剂盒建立在依赖于ATP 的萤火虫萤光素酶反应上，该反应可产生光信号。ATP 先与Pgp 共同孵育，然后Pgp ATP 酶反应被终止, 未被消耗的 ATP 由萤光素产生的发光信号而被检测出来。Pgp依赖的发光信号的降低反映了Pgp 对ATP 的消耗，因此信号减少得越多，Pgp 的活力就越高。相应地，若待测样品中含有可刺激Pgp ATP 酶的化合物，这个反应产生的发光信号会显著低于未经处理的对照组。
The Pgp-Glo™ Assay Systems provide the necessary reagents for performing luminescent P-glycoprotein (Pgp) ATPase assays. Pgp, also known as MDR1 and ABCB1, is a 170kDa integral plasma membrane protein that functions as an ATP-dependent drug efflux pump and plays an important role in multidrug resistance and certain adverse drug-drug interactions. Compounds that interact with Pgp can be identified as stimulators or inhibitors of its ATPase activity. Compounds that are substrates for transport by Pgp typically stimulate its ATPase activity.
The Pgp-Glo™ Assay detects the effects of compounds on recombinant human Pgp in a cell membrane fraction. The assay relies on the ATP dependence of the light-generating reaction of firefly luciferase. ATP is first incubated with Pgp; then the Pgp ATPase reaction is stopped, and the remaining unmetabolized ATP is detected as a luciferase-generated luminescent signal. Pgp-dependent decreases in luminescence reflect ATP consumption by Pgp; thus the greater the decrease in signal, the higher the Pgp activity. Accordingly, samples containing compounds that stimulate the Pgp ATPase will have significantly lower signals than untreated samples.
Features - Benefits
Complete System:Cat.# V3591 includes all the reagents required to run the assay except the P-glycoprotein: A Pgp reaction buffer, MgATP, Verapamil, Na3VO4, and a lyophilized ATP detection reagent and its reconstitution buffer. Cat.# V3601 includes all the reagents provided in the Pgp-Glo™ System with the addition of Recombinant Human Pgp Membranes to provide a completely optimized kit.
Stable Activities:"Glow-type" signal allows processing of multiple samples without concern of variability over time.
Low False-Positive Rate:Use of a proprietary stabilized firefly luciferase and a proprietary luciferase assay formulation minimizes the incidence of false positives due to inhibition of luciferase by analytes when screening for compounds that affect Pgp activity.
Simple:The simple protocol makes the assay amenable to high-throughput screening in multiwell plates.
Applications
Screen drugs and new chemical entities for their capacity to modulate P-glycoprotein ATPase activity.
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