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With ATP as the phosphate donor, NAD+ kinase converts NAD+ to NADP+. NADP(H) supplies reducing power for biosynthesis and antioxidant defense. NAD+ kinase is an attractive target for antibacterial work.
Pack Sizes
Both sizes include the E. coli NAD+ kinase. The only difference is the number of assays: NADK100KE provides reagents for 100 assays and NADK500KE for 500 assays (384-well format), with component volumes scaled accordingly.
| 100 assays (NADK100KE) | 100 assays; enzyme included |
|---|---|
| 500 assays (NADK500KE) | 500 assays; enzyme included |
Kit Components
| Component | 100 assays (NADK100KE) | 500 assays (NADK500KE) |
|---|---|---|
| 10 x Assay buffer | 600 µl | 2 ml |
| 100 x NAD+ | 35 µl | 170 µl |
| 100 x E. coli NAD kinase | 35 µl | 170 µl |
| 100 x ATP | 35 µl | 170 µl |
| 100 x MUK Reagent A | 35 µl | 170 µl |
| 100 x MUK Reagent B | 35 µl | 170 µl |
| 10 x Fluorescence dye | 300 µl | 1.7 ml |
The 1 x buffer contains 50 mM Tris-HCl pH 8.0, 3 mM MgCl2, 0.2 mM EDTA, 0.5 mM DTT, 50 mM NaCl and 0.003% Brij-35.
Assay Principle
The kit measures the ADP formed in the kinase reaction (NAD+ + ATP -> NADP+ + ADP). MUK Reagents A and B and a fluorescence dye turn the ADP into a fluorescence signal read at Em 535 nm with excitation at 485 nm.
Specifications
| Assay Target | E. coli NAD+ kinase activity |
|---|---|
| Target Type | Kinase |
| Organism | E. coli |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Fluorescence |
| Detection Wavelength | Ex 485 nm / Em 535 nm |
| Plate Format | Black 384/96-well plate or cuvette |
| Instrument Required | Plate reader or fluorometer |
| Assay Time | About 62 min (2 min pre-incubation + 60 min reaction, then add dye and read) |
| Sizes | 100 assays, 500 assays |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped frozen. |
Protocol
The manufacturer's protocol describes the procedure and reaction volumes.
- E. coli NAD+ Kinase Assay (PDF, see Documents): Protocol for the E. coli NAD+ Kinase Assay Kit Plus.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this E. coli NAD+ kinase activity assay — including larger pack sizes, bulk reagents, bulk enzyme, or a protocol adapted to your samples. Click Talk to a Scientist, email support@biohippo.com, or explore our Research Services. Our team will be in touch shortly.