Recommended Dilution (serum & plasma):
Human: 8x / Mouse: 2x / Rat: 2x
This EIA kit is designed to detect human, mouse, and rat Cortisol.
Number of Targets Detected:
Compatible Sample Types:
- Cell Culture Supernatants
Method of Detection:
1, 2, or 5 x 96-Well Strip Microplate Kit
Cortisol is a naturally occurring pregnane corticosteroid and is also known as 11β,17α,21-trihydroxypregn-4-ene-3,20-dione. Cortisol is synthesized from cholesterol mainlyby the adrenal gland in the zona fasciculata. Release of Cortisolis increased in response to stress and low blood-glucose concentration. It functions to increase blood sugar through gluconeogenesis, to suppress the immune system, and to aid in the metabolism of fat, protein, and carbohydrates. It also decreases bone formation.
- Strip plates and additional reagents allow for use in multiple experiments
- Quantitative protein detection
- Establishes normal range
- The best products for confirmation of antibody array data
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Standard Peptide
- Assay Diluent(s)
- Biotinylated Peptide
- TMB One-Step Substrate
- Stop Solution
- Assay Diagram
- Positive Control Sample
- Capture Antibody
- User Manual
Other Materials Required
- Distilled or deionized water
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Orbital shaker
- Aluminum foil
- Saran Wrap
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- SigmaPlot software (or other software that can perform four-parameter logistic regression models)
- Prepare all reagents, samples and standards as instructed.
- Add 100 µl detection antibody to each well.
- Incubate 1.5 h at RT or O/N at 4°C.
- Add 100 µl standard or sample to each well.
- Incubate 2.5 h at RT.
- Add 100 µl prepared streptavidin solution.
- Incubate 45 min at RT.
- Add 100 µl TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl Stop Solution to each well.
- Read plate at 450 nm immediately.
Standard, biotinylated peptide, and positive control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened microplate wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.