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ADP is formed by ATP hydrolysis, including by kinases and other ATP-using enzymes. Measuring it helps check ATP stocks for ADP contamination, follow ATP hydrolysis, and track the activity of ADP-producing enzymes.
Pack Sizes
The only difference is the number of samples: MAD100K is enough for 100 samples and MAD500K for 500 samples in a standard black 384-well plate; see the Kit Components table for the volumes of each size.
| 100 assays (MAD100K) | 100 assays |
|---|---|
| 500 assays (MAD500K) | 500 assays |
Kit Components
| Component | 100 assays (MAD100K) | 500 assays (MAD500K) |
|---|---|---|
| 10 x Buffer | 350 µl | 1800 µl |
| 100 x MAD reagent 1 | 30 µl | 150 µl |
| 100 x MAD reagent 2 | 30 µl | 150 µl |
| 10 x Fluorescence dye | 300 µl | 1500 µl |
1 x Buffer: 50 mM Tris-HCl, pH 8.0, 3 mM MgCl2, 0.2 mM EDTA, 0.5 mM DTT, 50 mM NaCl, 0.003% Brij-35.
Assay Principle
Samples are mixed with a premix of buffer and two MAD reagents, and a fluorescence dye is then added; the signal (Ex 485 nm / Em 535 nm) is linear with ADP. ADP in unknown samples is calculated from a standard curve, Fc = a [ADP] + b, as [ADP] = (Fc - b) / a.
Specifications
| Assay Target | ADP concentration |
|---|---|
| Analyte | ADP |
| Analyte Category | Nucleotide |
| Assay Type | Concentration Assay |
| Detection Method | Fluorescence |
| Detection Wavelength | Ex 485 nm / Em 535 nm |
| Plate Format | 384/96-well plate or cuvette |
| Instrument Required | Plate reader or fluorometer |
| Assay Time | About 50 min (45 min reaction + 5 min detection) |
| Measuring Range | Sub-micromolar to low micromolar ADP |
| Selectivity | Very little background from UDP, GDP and CDP; ATP, UTP, GTP, CTP, AMP, UMP, GMP and CMP not detected |
| Compatibility | Compatible with common reaction buffers with magnesium; not compatible with samples with high DNA or RNA concentrations |
| 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.
- ADP assay (PDF, see Documents): Covers catalog # MAD100K, MAD500K.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this ADP concentration assay — including larger pack sizes, bulk reagents, 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.
