FT3
FT3 ELISA TEST KIT
Free Triiodothyronine 96 tests
1. Intended use
Immunoassay for the in vitro quantitative determination of free
triiodothyronine in human serum.
2. Summary
- Triiodothyronine is one of the thyroid hormones present in serum
which regulates metabolism. Determination of this hormone
concentration is important for the diagnostic differentiation of
euthyroid, hyperthyroid, and hypothyroid states. The major fraction
of total triiodothyronine is bound to the transport proteins (TBG,
prealbumin, albumin).
- Free triiodothyronine (fT3) is the physiologically active form of
the thyroid hormone triiodothyronine (T3).
- The determination of free T3 has the advantage of being independent
of changes in the concentrations and binding properties of the
binding proteins; additional determination of a binding parameter
(T-uptake, TBG) is therefore unnecessary. In normal thyroid
function, as the concentrations of the carrier proteins alter, the
total T3 level changes so that the FT3 concentration remains
constant.
- Thus,measurements of FT3 concentrations correlate more reliably
with clinical status than total T3 levels.
- or example, the increase in total T3 levels associated with
pregnancy, oral contraceptives and estrogen therapy result in
higher total T3 levels while the FT3 con- centration remains
basically unchanged. In addition, it has been found that the mean FT3 value has a
gradient decreasing from young to older.
- Reagents
3. Materials provided
• Coated Microplate, 8 x 12 strips, 96 wells, pre-coated with T3
derivant.
• Calibrators, 6 vials, 1 ml each, ready to use; Concentrations:
0(A), 2(B), 5(C), 10(D), 20(E) and 50(F) pmol/L.
• Enzyme Conjugate, 1 vial, 6.0 ml of HRP (horseradish peroxidase)
labeled sheep monoclonal Anti-T3 in Tris-NaCl buffer containing BSA
(bovine serum albumin). Contains 0.2% ProClin300 preservative.
• Substrate, 1 vial, 11ml, ready to use, (tetramethylbenzidine)
TMB.
• Stop Solution, 1 vial, 6.0 ml of 1 mol/l sulfuric acid.
• Wash Solution Concentrate, 1 vial, 25 ml (40X concentrated),
PBS-Tween wash solution.
• IFU, 1 copy.
• Plate Lid: 1 piece.
Materials required (but not provided)
• Microplate reader with 450nm and 620nm wavelength absorbent
capability.
• Microplate washer.
• Incubator.
• Plate shaker.
• Micropipettes and multichannel micropipettes delivering 50μl with
a precision of better than 1.5%.
• Absorbent paper.
• Distilled water.
4. Test Produce
Ensure the patients’ samples, calibrators, and controls are at
ambient temperature (18-25 ℃) before measurement. Mix all reagents
through gently inverting prior to use.
• Use only the number of wells required and format the microplates’
wells for each calibrator and sample to be assayed. • Add 50 µL of
calibrators or samples to each well.
• Add 50 µL of enzyme conjugate to each well.
• Shake the microplate gently for 30 seconds to mix.
• Cover the plate with a plate lid and incubate at 37 ℃ for 60
minutes.
• Discard the contents of the micro plate by decantation or
aspiration. If decanting, tap and blot the plate dry with absorbent
paper.
• Add 350 µL of wash solution, decant (tap and blot) or aspirate.
Repeat 4 additional times for a total of 5 washes. An automated
microplate strip washer can be used. At the end of washing, invert
the plate and tap out any residual wash solution onto absorbent
paper. • Add 100 µL of substrate to each well.
• Cover and incubate at ambient temperature (18-25℃)in the dark for
reaction for 20 minutes. Do not shake the plate after substate
addition. • Add 50 µL of stop solution to each well.
• Shake for 15-20 seconds to mix the liquid within the wells. It is
important to ensure that the blue color changes to yellow
completely. • Read the absorbance of each well at 450 nm ( using
620 to 630 nm as the reference wavelength to minimize well
imperfections) in a micro plate reader.The results should be read
within 30 minutes of adding the stop solution.
