Estradiol (E2) is an estrogen steroid hormone, also known as 17β-estradiol (to distinguish it from 17α-estradiol) or as estra-1,3,5(10)-triene-3,17β-diol. It is the major female sex hormone, and involved in the regulation of the estrous and menstrual female reproductive cycles. Estradiol is responsible for the female sexual development, and is important in the development and maintenance of female reproductive tissues. Estradiol is produced mainly by the granulosa cells of the ovaries, is also in other tissues including the testicles, the adrenal glands, fat, liver, the breasts, and the brain.

• 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
• Assay Diluent
• Lyophilized Standard
• Positive Control
• Capture Antibody
• HRP-conjugated secondary antibody
• Wash Buffer
• 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
• Distilled or deionized water
• Tubes to prepare standard or sample dilutions
• Orbital shaker
• Software which can perform four-parameter logistic regression models

1. Prepare all reagents, samples and standards as instructed.
2. Add 50 µL standards or samples to appropriate well. Incubate for 10 minutes at room temperature.
3. Add 50 µL Capture Antibody to appropriate well. Incubate 1 hour at room temperature or 37°C. And wash 5 times.
4. Add 100 µL prepared HRP-Secondary antibody solution. Incubate 1 hour at room temperature or 37ºC. And wash 5 times.
5. Add 100 µL TMB One-Step Substrate Reagent to each well. Incubate 5-20 minutes at room temperature or 37°C.
6. Add 50 µL Stop Solution to each well. Read at 450 nm immediately.

The entire kit may be stored at -20°C to -80°C for up to 1 year from the date of shipment. For extended storage, it is recommended to store it at -80°C. Avoid repeated freeze-thaw cycles.