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Guanylate kinase is essential for guanine nucleotide supply. It moves a phosphate from ATP to GMP, giving GDP and ADP, and is an attractive antibacterial target. This kit measures the ADP formed.
Pack Sizes
Both sizes include the E. coli guanylate kinase. The only difference is the number of assays: GMK100KE provides reagents for 100 assays and GMK500KE for 500 assays (384-well format), with component volumes scaled accordingly.
| 100 assays (GMK100KE) | 100 assays; enzyme included |
|---|---|
| 500 assays (GMK500KE) | 500 assays; enzyme included |
Kit Components
| Component | 100 assays (GMK100KE) | 500 assays (GMK500KE) |
|---|---|---|
| 10 x Assay buffer | 600 µl | 2000 µl |
| 100 x GMP | 35 µl | 170 µl |
| 100 x E. coli GMP 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 | 350 µl | 1700 µl |
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 (GMP + ATP -> GDP + 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 guanylate 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 Guanylate Kinase Assay (PDF, see Documents): Covers catalog # GMK100KE, GMK500KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this E. coli guanylate 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.