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
• Standard Peptide
• HRP-Conjugated Peptide
• Capture Antibody
• Positive Control
• Wash Buffer
• Assay Diluent(s)
• TMB One-Step Substrate Reagent
• Stop Solution

• Microplate reader capable of measuring absorbance at 450 nm
• 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
• Absorbent paper
• SigmaPlot software (or other software that can perform four-parameter logistic regression models)
• Tubes to prepare standard and sample dilutions
• Orbital shaker
• Distilled or deionized water
• Aluminum foil
• Plastic wrap

1. Prepare all reagents, samples and standards as instructed.
2. Add 100 µL detection antibody to each well. Incubate 1.5 hours at room temperature or overnight at 4ºC.
3. Add 100 µL standard or sample to each well. Incubate 2.5 hours at room temperature or overnight at 4ºC.
4. Add 100 µL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
5. Add 50 µL Stop Solution to each well. Read plate at 450 nm immediately.

The entire kit may be stored at -20°C to -80°C for up to 6 months from the date of shipment. For extended storage, it is recommended to store it at -80°C. Avoid repeated freeze-thaw cycles.