Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Tissue Homogenates (50 uL), EDTA Plasma (50 uL), Urine (10 uL)
Sensitivity
1.54 nmol/L
Assay Range
3.8 - 240 nmol/L (Cell Culture Supernates, Cell Lysates, Tissue Homogenates, EDTA Plasma, Urine)
Specificity
Cyclic AMP in mouse and rat samples
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Control Available
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QC53, Parameter Immunoassay Control Set 764 for Mouse/Rat cAMP -
Product Summary
The R&D Systems cAMP Immunoassay is a 3.5 hour competitive enzyme immunoassay designed to measure cAMP in cell culture supernates, cell lysates, tissue homogenates, plasma, and urine.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, Cell Lysates, Tissue Homogenates, EDTA Plasma, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 27.7 | 58.7 | 96.9 | 28.2 | 61.7 | 102 |
| Standard Deviation | 0.5 | 1 | 3.9 | 2.2 | 4.8 | 6.5 |
| CV% | 1.8 | 1.7 | 4 | 7.8 | 7.8 | 6.4 |
Recovery
The recovery of cAMP spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Samples (n=4) | 100 | 98-103 |
| Cell Lysates (n=2) | 106 | 106-107 |
| Mouse EDTA Plasma (n=4) | 97 | 92-101 |
| Rat EDTA Plasma (n=4) | 104 | 101-108 |
| Tissue Homogenates (n=4) | 110 | 105-115 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of cAMP were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.
Background: cAMP
Adenosine 3,5-cyclic monophosphate (cAMP) is one of the most important intracellular second messengers. Several hormones are known to activate cAMP and subsequent signaling cascades via their indirect influence on adenylate cyclase, the enzyme responsible for the conversion of ATP to cAMP.
Long Name
Cyclic AMP
Alternate Names
cAMP;
