Mouse Estradiol PharmaGenie ELISA Kit (SBRS0050)

1. Pre-Coated 96-well Strip Microplate
2. Wash Buffer
3. Standard Peptide
4. Assay Diluent(s)
5. Biotinylated Peptide
6. HRP-Streptavidin
7. TMB One-Step Substrate
8. Stop Solution
9. Assay Diagram
10. Positive Control Sample
11. Capture Antibody
12. Technical Manual

Other materials and equipment required:

The Assay Genie Mouse Estradiol PharmaGenie ELISA Kit (SBRS0050) will require other equipment and materials to carry out the assay. Please see list below for further details.

• Distilled or deionized water
• Precision pipettes to deliver 2 ul 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)

1. Prepare all reagents, samples and standards as instructed.
2. Add 100 ul detection antibody to each well.
3. Incubate 1.5 h at RT or O/N at 4°C.
4. Add 100 ul standard or sample to each well.
5. Incubate 2.5 h at RT.
6. Add 100 ul prepared streptavidin solution.
7. Incubate 45 min at RT.
8. Add 100 ul TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 ul Stop Solution to each well.
11. Read plate at 450 nm immediately.

Estradiol (E2) is an estrogen steroid hormone, also known as 17β-estradiol (to distinguish it from 17β-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.