NDP(ADP, GDP, UDP and CDP) assay Kits
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NDP(ADP, GDP, UDP and CDP) assay Kits
Introduction
NDP(ADP, GDP, UDP and CDP) are common products of NTP(ATP, GTP, UTP or CTP) hydrolysis in enzyme reactions or kinase reactions using NTP as the phosphate donor. The NDP assay kits are for measurements of 0.2 mM – 10 mM (micromolar) NDP concentrations. The assays are fluorescence-based and performed using regular black 96-well or 384-well plates or micro-cuvettes. Each assay kit contains the 10 x buffer, 100 x reagent 1, 100 x reagent 2 and 10 x dye. The assay reagents in kits MAD100K, MGD100K, MUD100K or MCD100K are sufficient for measurement of 100 samples using a standard black 384-well plate (Matrix 4318 plate). The assay reagents in kits MAD500K, MGD500K, MUD500K or MCD500K are sufficient for measurement of 500 samples using a standard black 384-well plate.
The kits can be used for measurement of NDP concentrations in an aqueous solution in the presence of NTP or NMP (AMP, GMP, UMP or CMP). The assay is not interfered by NTP or NMP. Contamination of NDP in the NTP or NMP samples is possible and will be detected by this NDP assay.
Protocol
The following protocol is for ADP assay. The protocols for GDP, UDP and CDP assays are similar to the ADP assay. The assay protocol is based on measurement using a standard black 384-well plate (Matrix 4318 plate). Adjust the volumes proportionally when more or less samples are measured.
Reagent preparation
1 x MAD buffer: Dilute the 10 x MAD buffer with water (10-fold dilution).
10 x MAD Reagent 1: Dilute the 100 x MAD reagent 1 with the water (10-fold dilution).
10 x MAD Reagent 2: Dilute the 100 x MAD reagent 2 with the 1x MAD buffer (10-fold dilution).
1 x Fluorescence dye: Dilute the 10 x Fluorescence dye with water (10-fold dilution).
ADP samples : Make 20 ml of ADP samples with concentrations from 0.1 mM to 10 mM.
1 x MAD 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
1. Mix 20 ul the ADP samples with 3 ul of the 10 x MAD buffer, 3 ul of 10 x MAD Reagent 1 and 3 ul of 10 x MAD Reagent 2 in the wells of a standard 384-well black plate for 30 min.
2. Add 30 ul of 1 x Fluorescence dye and read the fluorescence intensity at 535 nm with excitation at 485 nm.
Data
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Ordering Information
|
Catalog No. |
Product Name |
Size |
|
MAD100K |
MicroMolar ADP Assay kit |
100 assays |
|
MAD500K |
500 assays | |
|
MGD100K |
MicroMolar GDP assay kit |
100 assays |
|
MGD500K |
500 assays | |
|
MUD100K |
MicroMolar UDP assay kit |
100 assays |
|
MUD500K |
500 assays | |
|
MCD100K |
MicroMolar CDP assay kit |
100 assays |
|
MCD500K |
500 assays |
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