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Methylerythritol phosphate cytidyltransferase (IspD) is an enzyme of the non-mevalonate pathway of isoprenoid biosynthesis. This pathway is found in many pathogenic organisms and in plants but not in mammals, which makes IspD an attractive target for new antibiotics and herbicides.
IspD forms 4-diphosphocytidyl-2-C-methyl-D-erythritol from CTP and 2-C-methyl-D-erythritol 4-phosphate (MEP).
Pack Sizes
Both sizes include E. coli IspD and pyrophosphatase. The only difference is the number of assays: ISPD100KE provides reagents for 100 assays and ISPD500KE for 500 assays (384-well format), with component volumes scaled accordingly.
| 100 assays (ISPD100KE) | 100 assays; enzyme included |
|---|---|
| 500 assays (ISPD500KE) | 500 assays; enzyme included |
Kit Components
| Component | 100 assays (ISPD100KE) | 500 assays (ISPD500KE) |
|---|---|---|
| 10 x Buffer | 400 µl | 2 ml |
| 100 x MEP | 35 µl | 170 µl |
| 100 x CTP | 35 µl | 170 µl |
| 100 x E. coli IspD (200 nM) | 35 µl | 170 µl |
| 100 x Pyrophosphatase (PPase, 10 U/ml) | 35 µl | 170 µl |
| Phosphate detection dye (also sold separately as MicroMolar Phosphate Assay Reagent, BHT16300173) | 5 ml | 25 ml |
Assay Principle
The IspD reaction releases pyrophosphate, which pyrophosphatase converts into inorganic phosphate. A phosphate detection dye is then added and absorbance at 650 nm is read; the signal reflects IspD activity.
Specifications
| Assay Target | E. coli IspD activity |
|---|---|
| Target Type | Cytidyltransferase |
| Organism | E. coli |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Colorimetric |
| Detection Wavelength | Absorbance 650 nm |
| Plate Format | 384/96-well plate or cuvette |
| Instrument Required | Plate reader or spectrophotometer |
| Assay Time | About 65 min (60 min reaction + 5 min detection) |
| 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 IspD Assay (PDF, see Documents): Covers catalog # ISPD100KE, ISPD500KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this E. coli IspD 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.