www.immunometrics.co.uk web

Ethinyl Estradiol RIA kit

  • high quality
    - developed for clinical trials and pharmacokinetic studies
    - extraction RIA not subject to interference problems associated with direct serum/plasma immunoassays
  • high performance
    - detection limit 337 pmol/L (100 pg/mL)
    - between-assay CV better than 10% in the range 337-3370 pmol/L (100-1000 pg/mL)
  • convenient
    - all principal reagents are provided
    - uses inexpensive, robust equipment
    - each kit is sufficient for 100 assay tubes
    - shelf life of 1 year at 4-8°C
    - three internal Quality Control samples are provided

For information and prices please contact us.

Principle

The assay is an extraction competitive RIA for the quantitative measurement of Ethinyl Estradiol (EE2) in human serum or plasma. The EE2 extraction RIA is not subject to the well-documented interference problems associated with direct serum/plasma immunoassays. Ethinyl estradiol in the sample is extracted with diethyl ether. The extract is dried and redissolved in Assay Buffer. The redissolved extract is assayed for ethinyl estradiol by 3H-RIA (tritium labelled radioimmunoassay with charcoal separation). The samples and standards are incubated with ethinyl estradiol, tritium tracer and antibody. Separation of free ethinyl estradiol from antibody-bound ethinyl estradiol is achieved by means of a charcoal separation reagent. The assay has five main stages.

  • Ethinyl estradiol in the sample (150 µL) is extracted with diethyl ether. The extract is dried and redissolved in Assay Buffer. Ethinyl estradiol standards are prepared in Assay Buffer.
  • Anti-ethinyl estradiol antibody and tritium labelled ethinyl estradiol tracer are incubated with standards and samples for 18-24 hours at 4°C.
  • A charcoal separation reagent is added, and incubated for 30-35 minutes at 4°C., followed by centrifugation at 4°C for 5 minutes.
  • After centrifugation, the supernatants are transferred to tubes containing scintillation cocktail.
  • The tubes are then placed in a Beta (liquid scintillation) Counter. The radioactive count of each tube can be measured and the results calculated using a data processing program.
Kit Contains

EE2 RIA Standard
EE2 RIA Antiserum
EE2 RIA Tritium Label
EE2 RIA Charcoal Reagent
EE2 RIA Dextran Reagent
EE2 RIA Gelatin Reagent
EE2 RIA Internal QC Samples (Low, Medium and High)
Instruction booklet

Reagents Required

Distilled/deionised water, Scintillation Cocktail, Diethyl ether, Sodium dihydrogen phosphate, Disodium hydrogen phosphate, Sodium chloride, Thimerosal. EQUIPMENT REQUIRED Glass test tubes, Pipettes, Vortex mixer, Magnetic stirrer, Centrifuge, Water bath, and Beta (liquid scintillation) counter.


Immunometrics (UK) Ltd, 280 Munster Road, London, SW6 6BQ, UK. Tel: + 44 + (0) 20 7386 9636