• high performance

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  detection limit 20 pmol/L (5 pg/mL)

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  within-assay CV below 15% between 0.1-11 nmol/L (25-3000pg/mL)

• low cost

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  all reagents are provided - only test tubes and water are required

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  uses inexpensive, robust equipment

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  each kit is sufficient for 100 assay tubes

• convenient

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  direct serum assay (no pre-extraction of samples required)

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  magnetic separation

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  shelf life of 9 months at 4-8°C

For information and prices please contact us.

The assay is a direct competitive EIA for the quantitative measurement of Estradiol in human serum or plasma. A specific agent displaces Estradiol from binding proteins, making it available for antibody binding. Estradiol then reacts successively with an anti-Estradiol antibody linked to fluorescein isothiocyanate (FITC) and alkaline phosphatase labelled Estradiol . An anti-FITC antibody bound to magnetic particles then separates the estradiol/estradiol label-antibody complex from unbound components. A further incubation with substrate produces colour in inverse proportion to the amount of estradiol present. The assay has five main stages:

• Displacing agent and anti-estradiol antibody conjugated to FITC are incubated with sample (150 µL) for 20 minutes at 37°C.
• Alkaline phophatase labelled estradiol tracer is added and the incubation continued for a further 20 minutes at 37°C.
• Anti-FITC antibody coupled to magnetic particles is added and incubated for 5 minutes at 37°C. Anti-estradiol antibody is then isolated by means of a magnetic wash step.
• A coloured enzyme substrate is incubated with the particles for 30 minutes at 37°C. The presence of alkaline phosphatase causes a colour change from yellow to pink. The reaction is terminated by addition of Stop Solution.
• The tubes are then placed in the spectrophotometer or colorimeter. The optical density (at 550nm) of each tube can be measured and the results calculated using a data processing program.

Incubation times	20 mins + 20 mins + 5 mins + 30 mins
Standard Range	0-11 nmol/L 0-3000 pg/mL
Normal Ranges	Male <0.22 nmol/L Female: Follicular phase 0.04-0.18 nmol/L Female: Pre-ovulatory peak 0.56-0.87 nmol/L Female: Luteal phase 0.18-0.63 nmol/L Female: Postmenopausal <0.24 nmol/L
Detection Limit	<0.018   nmol/L
Assay Reproducibility	Within-batch ±1SD nmol/L %CV 0.06±0.01 12.6 0.7±0.03 3.9 2.5±0.1 4.4 6.3±0.3 4.9 10.8±0.8 7.7
Assay Reproducibility	Between-batch ±1SD nmol/L %CV 0.08±0.01 13.0 0.8±0.1 6.3 2.4±0.1 5.9 5.9±0.7 11.9 9.1±0.8 9.2

Estradiol EIA Standards Estradiol EIA Antiserum Estradiol EIA Separation Reagent Estradiol EIA Enzyme Label Estradiol EIA Wash Concentrate Estradiol EIA Serum Diluent

EIA Substrate Reagent EIA Substrate Buffer EIA Stop Buffer Estradiol EIA Internal QC Sample Instruction booklet

Test tubes, pipettes, multivortex mixer, magnetic separator, 37°C water- bath and a colorimeter or spectrophometer.