Kinase Selectivity Profiling System TK-4 + ADP-Glo?

• 产品编号：V6923      品牌：promega       原厂货号：V6923
• 产品分类：蛋白质检测与分析 > 酶检测和活性分析 > 蛋白酶/肽酶检测
• 应用分类：

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描述：

The Kinase Selectivity Profiling Systems are easy-to-use kits for performing kinase selectivity profiling that rely on the ADP-Glo™ Kinase Assay technology (Figure 1). Each system includes kinase and substrate pairs organized in an easy-to-use 8-tube strip format optimized for fast and simple kinase profiling reactions (see Figures and Tables tab for complete listing of enzymes and substrates). Kinase Selectivity Profiling Systems offer kinases grouped either in single kinase family strips or as a general panel of kinases representative of the human kinome for a broad kinase profile. Each profiling system contains the reagents needed to complete a profile for a compound, including kinase reaction buffer, eight kinases in each multiwell strip and eight corresponding substrates and cofactors in another multiwell strip (Figure 2). The General Panel contains 24 kinases arranged in three 8-well strips. The kinase stock solutions are standardized in a way that, when kinases are diluted to the final concentration in the kinase reaction, the kinase activity will result in optimal ATP to ADP conversion in 5μl reactions (384-well plate), with a signal-to-background ratio of more than ten when used in conjunction with the ADP-Glo™ Kinase Assay (1). The substrate stock solutions are standardized in a similar fashion and provided in a second 8-tube strip with the substrates at corresponding positions.

ADP-Glo™ Kinase Assay is a luminescent kinase assay that measures ADP formed from a kinase reaction; ADP is converted into ATP, which is converted into light by Ultra-Glo™ Luciferase. The luminescent signal positively correlates with ADP amount and kinase activity. The assay is well suited for measuring the effects chemical compounds have on the activity of a broad range of purified kinases—making it ideal for both primary screening as well as kinase selectivity profiling. The ADP-Glo™ Kinase Assay can be used to monitor the activity of virtually any ADP-generating enzyme (e.g., kinase or ATPase) using up to 1mM ATP.

Features - Benefits

• Fast Turnaround Time: Lead compounds can be profiled in-house in a matter of hours versus days when compounds are sent out.
• Flexible Kinase Inhibitor Profiling: Each system has enough material to profile up to twenty compounds at a single dose or create a dose-response for two compounds against the eight kinases at once.
• Fast and Simple Reaction Assembly: Two quick dilutions provide working stocks of kinases and substrate/cofactor solutions.
• Optimized Kinase Activity for Inhibitor Profiling: All kinases have been optimized to provide 10–30% ADP production when assayed at 10µM ATP.
• Formatted Strips Provide Access to Eight Kinases at One Time: Kinases from singular kinase families are grouped together for a more relevant selectivity profile.
• Accurate: Accurately measure ADP levels at a wide range of starting ATP concentrations; activity measured truly reflects kinase activity and produces accurate IC₅₀ values comparable to radioactivity-based assays.
• Stable Luminescent Signal: Perform batch plate processing without need for strictly timed incubations; flexible.

Applications

• Selectivity screening of library compounds for effects on target kinases.
• Profile hits and leads for their potency against families of related kinases.

References

1. Zegzouti, H. et al. (2009) ADP-Glo: A bioluminescent and homogeneous ADP monitoring assay for kinases. Assay Drug Dev. Technol. 7, 560–72.

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