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Immunometrics:
T3 EIA kit

  • high performance
    - detection limit 0.23 nmol/L (0.15 ng/mL)
    - within-assay CV less than 15% between 0.2 - 15 nmol/L ( 0.13 - 10 ng/mL)
  • low cost
    - all reagents are provided - only test tubes and water are required
    - uses inexpensive, robust equipment
    - each kit is sufficient for 100 assay tubes
  • convenient
    - direct serum assay
    - magnetic separation
    - shelf life of 9 months at 4-8°C

For information and prices please contact us.

Principle

The assay is a direct competitive EIA for the quantitative measurement of Total Triiodothyronine (Total T3) in human serum or plasma. A specific agent displaces T3 from binding proteins, making it available for antibody binding. T3 then equilibrates with fluorescein labelled T3 (T3 Derivative) in binding to a limited amount of alkaline phosphatase labelled anti-T3 antibody. An anti-fluorescein antibody bound to magnetic particles then separates the T3/fluorescein T3-antibody complex from unbound components. A further incubation with substrate produces colour in inverse proportion to the amount of T3 present. The assay has four main stages:

  • Displacing agent, alkaline phosphatase labelled anti-T3 antibody, and fluorescein-T3 derivative are incubated with sample (100 µL) for 15 minutes at 37°C.
  • Anti-fluorescein antibody coupled to magnetic particles is added and incubated for 10 minutes at 37°C. Anti-T3 antibody is then isolated by means of a magnetic wash step.
  • A coloured enzyme substrate is incubated with the particles for 15 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 or 492nm) of each tube can be measured and the results calculated using a data processing program.

See diagram popup.

Kit Parameters

Incubation times 15 mins + 10 mins + 15 mins
Standard Range 0.15.4 nmol/L
0-10 ng/mL
Normal Ranges
Adults 1.21-2.66 nmol/L
Pregnant: 2nd Trimester 1.63-3.34 nmol/L
Pregnant: 3rd Trimester 1.18-3.57 nmol/L
Detection Limit <0.23   nmol/L
Assay Reproducibility
Within-batch ±1SD
nmol/L %CV
1.8±0.06 3.50
4.8±0.1 2.7
9.7±0.3 3.0
Assay Reproducibility
Between-batch ±1SD
nmol/L %CV
2±0.1 4.4
5±0.2 4.0
9.7±0.5 4.6

Kit Contains

T3 EIA Standards
T3 EIA Antiserum
T3 EIA Separation Reagent
T3 EIA Derivative
T3 EIA Wash Concentrate
EIA Substrate Reagent
EIA Substrate Buffer
EIA Stop Buffer
T3 EIA Internal QC Sample
Instruction booklet

Equipment Required

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

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