FT4 ELISA TEST KIT Free Thyroxine
1. Intended use
Immunoassay for the in vitro quantitative determination of free
thyroxine in human serum.
2. Summary (1,2 3, 4)
The thyroid hormone thyroxine (T4) is physiologically part of the
regulating circuit of the thyroid gland and has an effect on
general metabolism. The major fraction of the total thyroxine is
bound to transport proteins (TBG, prealbumin, and albumin). The
free thyroxine (fT4) is the physiologically active thyroxine
component.
The determination of free thyroxine is an important element in
clinical routine diagnostics. Free T4 is measured together with TSH
when thyroid function disorders are suspected. The determination of
fT4 is also suitable for monitoring thyrosuppressive therapy.
The determination of free T4 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.
3. Test principle
Competition principle. Total duration of assay: 80 minutes.
• Sample, T4 derivant coated microwells and enzyme labeled Anti-T4
are combined.
• During the incubation, T4 derivant coated on microwells and FT4
present in the sample compete for binding to the enzyme labeled
antibodies.
• After washing, a complex is generated between the solid phase and
enzyme[1]linked antibodies by immunological reactions.
• Substrate solution is then added and catalyzed by this complex,
resulting in a chromogenic reaction. The resulting chromogenic
reaction is measured as absorbance.
• The color intensity is inversely proportional to the amount of
FT4 in the sample
4.Reagents
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), 5(B), 10(C),20(D), 50(E) and 100(F) pmol/L.
• Enzyme Conjugate, 1 vial, 6.0 ml of HRP (horseradish peroxidase)
labeled mouse monoclonal Anti-T4 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.
5. 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
