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The dipeptide D-Ala-D-Ala is a building block for bacterial peptidoglycan. D-Alanine:D-Alanine ligase (Ddl) makes it by joining two D-Alanine molecules, and this step is linked to ATP hydrolysis, which releases ADP and inorganic phosphate.
Pack Sizes
Both pack sizes include the E. coli D-Ala:D-Ala ligase. The only difference is the number of assays: DDA100KE provides reagents for 100 assays and DDA500KE for 500 assays (384-well format), with component volumes scaled accordingly.
| 100 assays (DDA100KE) | 100 assays; enzyme included |
|---|---|
| 500 assays (DDA500KE) | 500 assays; enzyme included |
Kit Components
| Component | 100 assays (DDA100KE) | 500 assays (DDA500KE) |
|---|---|---|
| 10 x Buffer | 400 µl | 2 ml |
| 100 x Enzyme Substrate | 35 µl | 170 µl |
| 100 x E. coli D-Ala:D-Ala ligase (200 nM) | 35 µl | 170 µl |
| Phosphate detection dye (also sold separately as MicroMolar Phosphate Assay Reagent, BHT16300173) | 5 ml | 25 ml |
Assay Principle
The kit measures the inorganic phosphate generated by the ligation reaction with a phosphate detection dye; absorbance at 650 nm is proportional to ligase activity, so inhibitors reduce the signal.
Specifications
| Assay Target | E. coli D-Ala:D-Ala ligase activity |
|---|---|
| Target Type | Ligase |
| 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 D-Alanine D-Alanine Ligase Assay (PDF, see Documents): Covers catalog # DDA100KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this E. coli D-Ala:D-Ala ligase 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.