Cytidine Deaminase Assay Kit (ab239723)
Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Tissue
Overview
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Product name
Cytidine Deaminase Assay Kit -
Detection method
Fluorescent -
Sample type
Tissue -
Product overview
Cytidine Deaminase Assay Kit (ab239723) uses Cytidine Deaminase (CDA) to convert cytidine to uridine and NH3, as intermediates. The intermediate products then react with a proprietary Reaction Mix to generate a stable fluorophore that can be detected fluorometrically (Ex/Em= 410/470 nm). The kit is suitable for measuring CDA activity as low as 1 µUnit (1 pmole/min/well) in biological samples. The kit allows for the evaluation of CDA activity for comparison of activity rates in normal versus tumor tissues. Samples types include animal tissues such as mouse or rat spleen, liver and kidney.
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Platform
Microplate reader
Properties
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Storage instructions
Please refer to protocols. -
Components 100 tests Ammonium Chloride Standard 1 x 0.1ml CDA Assay Buffer 1 x 25ml CDA Substrate 1 vial Cytidine Deaminase (CDA) 1 vial Developer A 1 x 1.2ml Microplate Sealing Film 1 unit -
Relevance
Cytidine Deaminase (3.5.4.5) is a homotetramer enzyme that catalyzes the irreversible deamination of cytidine to produce uridine, thus maintaining the intracellular pyrimidine pool of uridine. Cytidine as well as adenosine, thymidine, guanosine and uridine are one of the five standard nucleosides with which nucleic acids are composed. Nucleosides accumulate or are recycled from salvage pathways within the cell and play an important role in DNA and RNA synthesis and subsequent cell division. Tumorigenesis is intricately linked to metabolism of macromolecular precursors, specifically, the building blocks of DNA and RNA. Thus tumor cells have greater concentrations of uridine and cytidine analogues (402 µM +/- 252) compared to normal cells (83 µM +/- 133), to support the increased demand for nucleotides in mitosis in actively dividing cells.
Images
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Ammonium Chloride Standard Curve.
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Estimation of Cytidine Deaminase Activity in mouse (26 µg) and rat (20 µg) kidney tissues.Mouse and rat kidney tissues were homogenized in CDA Assay Buffer with a dounce homogenizer. The supernatant was filtered three times with a 10 kD filter. Protein concentration was determined with a BCA Assay Protein Quantitation Kit and CDA Activity determined following the kit protocol. For each sample, 2.5 µL of washed supernatant diluted 1/10 was analyzed. Data are an average of 4 replicates, each analyzed in two separate experiments.
