Transcreener® CD73 Activity Assay
The CD73 activity assay for HTS and hit-to-lead. Measure specific activity of CD73 in a simple mix-and-read format. The kit is designed for compound library screening and follow-up inhibitor discovery.
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The Transcreener CD73 Assay is a far-red, competitive fluorescence polarization (FP) assay. The biochemical activity assay is designed to be used with enzymes such as CD73 that produce the product adenosine (ADO). Based on the proven Transcreener ADP2 Assay, the assay uses a coupling enzyme to convert ADO into ADP which is then can be measured with a reliable FP readout.
Also known as ecto-5' nucleotidase or 5'-nucleotidase, CD73 is an anchored cell surface protein that plays a critical role in purinergic signaling by catalyzing the hydrolysis of AMP into ADO and phosphate. Multiple lines of research have shown that adenosine is immunosuppressive in the tumor microenvironment, and ectonucleotidase enzymes have emerged as promising immuno-oncology targets.
The Transcreener assay is designed specifically for high throughput screening (HTS), with a single addition, mix-and-read format. It offers reagent stability and compatibility with commonly used multimode plate readers.
The Transcreener CD73 FP Assay provides the following benefits:
- A simple single addition CD73 activity assay
- Detection of unlabeled adenosine
- Easy to use, homogenous, one-step format
- Robust Assay Z€™ > 0.7 under initial velocity conditions
- Far-red fluorescent readout minimizes compound interference
- A safe, non-radioactive method
Applications:
- Measure Enzymatic Activity of CD73
- Screen Compound Libraries for CD73 Inhibitor
- Quantify Inhibitor Potency (IC50)
- Inhibitor Selectivity Profiling
The DNA damage response and innate immunity
Innate immunity and DNA repair are critical sense-response systems that have evolved to protect cells from external and internal threats: tumors, invading pathogens, toxic agents, and physical wounds. Both systems are the focus of intense research and drug discovery activity to enable more effective therapies for cancer and debilitating autoimmune and inflammatory diseases. Interconnections between innate immune responses triggered by nucleic acids and DNA repair pathways have emerged, and they are rapidly being translated into exciting, new therapeutic strategies. In this application note, explore how BellBrook Labs are trying to accelerate these efforts, showing biochemical assays for key enzymes in the innate immunity and DNA damage response (DDR) pathways using the Transcreener® HTS Assay platform.
