Aldosterone Parameter Assay Kit
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Summary
Precision
Recovery
Linearity
Product Datasheets
Preparation and Storage
Related Research Areas
Aldosterone Parameter Assay Kit Summary
Format
96-well strip plate
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (34 uL), Serum (34 uL), EDTA Plasma (34 uL), Heparin Plasma (34 uL), Urine (10 uL)
Sensitivity
22.4 pg/mL
Assay Range
24.7 - 6,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Specificity
Measures Aldosterone levels in various 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.
Product Summary
The Parameter Aldosterone assay is a 3.5 hour competitive enzyme immunoassay designed to measure Aldosterone in cell culture supernates, serum, plasma, and urine.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of kit components
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean | 278 | 878 | 2536 | 263 | 832 | 2419 |
| Standard Deviation | 9.49 | 17.3 | 34.4 | 31.7 | 83 | 230 |
| CV% | 3.4 | 2 | 1.4 | 12.1 | 10 | 9.5 |
Recovery
The recovery of Aldosterone spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 94 | 89-99 |
| EDTA Plasma (n=4) | 89 | 76-110 |
| Heparin Plasma (n=4) | 89 | 79-107 |
| Serum (n=4) | 88 | 78-108 |
| Urine (n=4) | 101 | 95-106 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Aldosterone were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay as directed in the Sample Preparation section.
Product Datasheets
Preparation and Storage
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.
Assay Procedure
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Aldosterone Primary Antibody Solution to each well (excluding the NSB wells). Cover with a plate sealer, and incubate at room temperature for 1 hour on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 50 µL of Assay Diluent RD1-120 to each well.
- Add 75 µL of Standard, control, or sample to the appropriate wells.
- Add 75 µL of Calibrator Diluent RD5-69 to the zero standard (B0) wells and NSB wells.
- Add 50 µL of Aldosterone Conjugate to all wells. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Aldosterone Primary Antibody Solution
50 µL Assay Diluent RD1-120
75 µL Standard, Control, or Sample
75 µL Calibrator Diluent RD5-69
50 µL Aldosterone Conjugate
200 µL Substrate Solution
100 µL Stop Solution
FAQs
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