E. coli D-Alanine:D-Alanine Ligase Assay Kit Plus

SKU:BHT16300034
Overview
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Colorimetric E. coli D-Alanine: D-Alanine Ligase Assay Kit Plus for measuring Ddl activity and screening antibacterial inhibitors. The released phosphate is read at 650 nm in 384-well or 96-well plates or a cuvette. Enzyme included; available in 100-assay and 500-assay sizes.
Assay Target E. coli D-Ala:D-Ala ligase activity
Detection Method Colorimetric
Detection Wavelength Absorbance 650 nm
Plate Format 384/96-well plate or cuvette
Assay Time About 65 min
Options selector
Catalog no. Size
DDA100KE 100 assays
DDA500KE 500 assays
Available Options

Select the variant that best fits your experiment. Availability and lead time may vary by option.

  • Options: Size: 100 assays, 500 assays
  • Lead time: options listed in "Availability Content"; other statuses may take longer.
  • Storage: Please contact us to confirm storage conditions.
  • Shipping: Shipped frozen.
  • Upon receipt: store at recommended temperature as soon as possible.
  • Sales terms and conditions: Please review prior to ordering.
Field Specification
Organism E. coli
Applications
  • Enzyme Activity
  • Compound Profiling
Assay type
  • Enzyme Activity Assay
  • Inhibitor Screening Assay
Assay target E. coli D-Ala:D-Ala ligase activity
Assay area Antibacterial Drug Discovery
Target type Ligase
Detection method
  • Colorimetric
Detection wavelength Absorbance 650 nm
Instrument required Plate reader or spectrophotometer
Plate format 384/96-well plate or cuvette
Assay time
  • About 65 min
Storage Please contact us to confirm storage conditions.
Shipping Shipped frozen.
Catalog no. (Mfr.) DDA100KE | DDA500KE
Main SKU BHT16300034
Drug Target Assay Kit

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

For Research Use Only (RUO). Not for use in diagnostic or therapeutic procedures. The E. coli D-Alanine:D-Alanine Ligase Assay Kit Plus is shipped frozen. Follow standard laboratory safety practices when handling the reagents. Please contact us to confirm storage conditions.
Q.What does this kit measure?
A.It measures the activity of E. coli D-Alanine:D-Alanine ligase by detecting the inorganic phosphate released when the enzyme joins two D-Alanine molecules and hydrolyzes ATP. It is used to screen inhibitors of this bacterial cell wall enzyme.
Q.What is the difference between the options?
A.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.
Q.Is a protocol available?
A.Yes. The manufacturer's protocol (E. coli D-Alanine D-Alanine Ligase Assay, linked under Documents) covers catalog # DDA100KE.
Q.How long does the assay take?
A.About 65 min (60 min reaction + 5 min detection). See the protocol for the exact steps.
Q.How should the kit be stored?
A.The E. coli D-Alanine:D-Alanine Ligase Assay Kit Plus is shipped frozen. Please contact us for long-term storage conditions.

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Experience the power of Celltrypse™, c-LEcta's innovative enzyme solution for gentle and efficient cell dissociation. Request your free sample and discover a superior alternative for your cell culture workflows.

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