MicroELISA

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The EIAgen Free T3 Kit is an EIA kit for the quantitative determination of free triiodothyronine (fT3) in human serum.

Triiodothyronine (T3), a thyroid hormone, circulates in blood almost completely bound (>99.5%) to carrier proteins (1,2). The main transport protein is thyroxine-binding globulin (TBG). However, only the free (unbound) portion of triiodothyronine is believed to be responsible for the biological action. Further, the concentrations of the carrier proteins are altered in many clinical conditions, such as pregnancy. In normal thyroid function as the concentrations of the carrier proteins alters, the total triiodothyronine concentration remains constant. Thus, measurements of the free triiodothyronine concentrations correlate more reliably with clinical status than total triiodothyronine levels.

For example, the increase in total T3 levels associated with pregnancy, oral contraceptives and estrogen therapy result in higher total T3 levels while the free T3 concentration remains basically unchanged.

Competitive Enzyme Immunoassay – Analog Method for Free T3

The essential reagents required for a solid phase enzyme immunoassay include antibody anti-T3 coated on microwells, enzyme-T3 conjugate and native free T3 antigen. The enzyme-T3 conjugate should have no measurable binding to serum proteins especially TBG and albumin. The method achieves this goal.

Upon mixing, in the coated microwells, the enzyme-T3 conjugate and a serum containing the native free T3 antigen, a competition reaction results between the native free T3 and the enzyme-T3 conjugate for a limited number of insolubilized binding sites.

After equilibrium is attained, the antibody-bound fraction is separated from unbound antigen by decantation or aspiration. The enzyme activity in the antibody-bound fraction is inversely proportional to the native free antigen concentration.

The activity of the enzyme present on the surface of well is quantitated by reaction with a substrate to produce colour.

By utilizing several different calibrators of known antigen concentration, a dose response curve can be generated from which the antigen concentration of an unknown can be ascertained.

The kit contains reagents for 96 tests (code LI4039).

Bring reagents to room temperature before use.