Aldosterone Parameter Assay Kit Summary
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine
|Intra-Assay Precision||Inter-Assay Precision|
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|
Preparation and Storage
- 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.
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