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GDP is formed by GTP hydrolysis, for example by GTPases. This kit measures GDP from sub-micromolar to low micromolar levels, for example to check GDP contamination in GTP samples, to follow GTP hydrolysis or to measure GDP-forming enzyme activities.
Pack Sizes
Both sizes contain the same reagents. The only difference is the number of assays: MGD100K is for 100 samples and MGD500K for 500 samples in 384-well plates, with component volumes scaled accordingly.
| 100 assays (MGD100K) | 100 assays |
|---|---|
| 500 assays (MGD500K) | 500 assays |
Kit Components
| Component | 100 assays (MGD100K) | 500 assays (MGD500K) |
|---|---|---|
| 10 x Buffer | 350 µl | 1800 µl |
| 100 x MGD Reagent 1 | 30 µl | 150 µl |
| 100 x MGD Reagent 2 | 30 µl | 150 µl |
| 10 x Fluorescence dye | 300 µl | 1500 µl |
The 1 x Buffer is 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
GDP samples are mixed with a premix of buffer and MGD Reagents 1 and 2, then a fluorescence dye is added and the signal is read at 535 nm (excitation 485 nm). ADP is also detected, UDP and CDP give very little background, and nucleoside triphosphates and monophosphates are not detected.
Specifications
| Assay Target | GDP concentration |
|---|---|
| Analyte | GDP |
| 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 micro-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 GDP |
| Selectivity | ADP also detected; UDP and CDP very low background; NTPs and NMPs not detected |
| Compatibility | Compatible with common reaction buffers with magnesium; not compatible with samples containing ADP or high DNA or RNA levels |
| 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.
- GDP assay (PDF, see Documents): Covers catalog # MGD100K, MGD500K.
Safety & Handling
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