NAD⁺ Metabolism Enzyme
Combo Assay Kit
Traditional NAD⁺ assays measure only metabolite levels, offering limited insight into cellular function.
The Signosis NAD⁺ Metabolism Enzyme Combo Assay Kit combines NAD⁺ quantification with functional measurement of SIRT, NAMPT, and PARP activity, providing a broader view of NAD⁺ metabolism in a single kit.
By examining NAD⁺ availability together with pathways involved in its biosynthesis, utilization, and signaling, researchers can better investigate changes associated with metabolism, aging, DNA repair, cellular stress, and drug response.
Key Measurements
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NAD⁺ Level – Quantifies cellular NAD⁺ availability.
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SIRT Activity – Measures activity of NAD⁺-dependent sirtuin enzymes involved in metabolic regulation and cellular stress responses.
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NAMPT Activity – Evaluates a key rate-limiting enzyme in the NAD⁺ salvage pathway.
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PARP Activity – Measures NAD⁺-dependent ADP-ribosylation associated with DNA damage and repair.
Principle
The NAD⁺ Enzyme Combo Assay Kit can measure NAD⁺ levels and the activity of two key NAD⁺-dependent enzymes: SIRT and NAMPT. Each marker is detected using a distinct and validated method:
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NAD⁺ is measured by first reducing NAD⁺ to NADH with alcohol dehydrogenase, and then measuring the NADH with WST-8, which is reduced by NADH to form a yellow color that can be measured at an absorbance of 450 nm.
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SIRT is detected using a FRET-based peptide substrate, which is deacetylated and cleaved by a SIRT/protease combination, and produces fluorescence after being released from the quencher.
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NAMPT is detected through a series of enzyme reactions. NAMPT first converts NAM into NMN. NMN is then converted by NMNAT into NAD⁺. Lastly, NAD⁺ is reduced by alcohol dehydrogenase into NADH, which is measured colorimetrically with WST-8.
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PARP is detected through its ability to perform ADP-ribosylation with a biotin-tagged substrate. PARP first attaches a biotin-tagged substrate to a plate coated with histones. Then, streptavidin-HRP and an HRP substrate is used to detect the biotin-tagged substrate bound to the plate. The signal is measured colorimetrically at an absorbance of 450 nm.
Products
Analyte | Product | SKU | Price (USD$) |
|---|---|---|---|
Combo | EA-7064 | 1800 | |
SIRT | EA-7061 | 650 | |
NAMPT | EA-7062 | 650 | |
PARP | EA-7063 | 550 | |
NAD⁺ | EA-7018 | 300 |
Benefits
Comprehensive Analysis
Assess both NAD⁺ levels and the activity of critical NAD⁺-dependent enzymes with one kit.
Convenient and Efficient
Combines multiple assays into a single platform to streamline experimental workflows.
Versatile Research Tool
Applicable to a wide range of biological studies, from basic cellular research to disease modeling.
High Sensitivity and Specificity
Designed to deliver accurate and reproducible measurements in cell or tissue samples.
