Transcreener^® CD73 Activity Assay by BellBrook Labs

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 Inhibitors
• Quantify Inhibitor Potency (IC50)
• Inhibitor Selectivity Profiling