MicroELISA

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EIAgen LH kit is solid phase enzyme immunoassay for the quantitative determination of Luteinizing Hormone (LH) in human serum.

Luteinizing hormone (LH) is a glycoprotein consisting of two subunits with a molecular mass of 30,000 daltons. The α-subunit is similar to other pituitary hormones [follicle stimulating hormone (FSH), thyroid stimulating hormone (TSH) and chorionic gonadotropin (HCG)] while the b-subunit is unique. The b-subunit confers the biological activity to the molecule.

The α -subunit consists of 89 amino acid residues while the b-subunit contains 129 amino acids. The carbohydrate content is between 15% and 30%.

The clinical usefulness of the measurement of luteinizing hormone (LH) in ascertaining the homeostasis of fertility regulation via the hypothalamic - pituitary - gonadal axis has been well established (1,2). In addition, the advent of in vitro fertilization (IVF) technology to overcome infertility-associated problems has provided the impetus for rapid improvement in LH assay methodology from the technically demanding bioassay (3) to the procedurally simple and rapid immunoenzymometric assays.

Immunoenzymometric assay.

In this method, LH calibrators, patient specimens and/or controls (containing the native antigen) are first added to streptavidin coated wells. Biotinylated monoclonal and enzyme labeled antibodies are added and the reactants mixed: these antibodies have high affinity and specificity and are directed against distinct and different epitopes of LH.

Reaction between the various LH antibodies and native LH occurs in the microwells without competition or steric hindrance forming a soluble sandwich complex.

Simultaneously, the complex is deposited to the well through the high affinity reaction of streptavidin and biotinylated antibody.

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 directly proportional to the native antigen concentration. The activity of the enzyme present on the surface of the well is quantitated by reaction with a suitable substrate to produce colour. By utilizing several different calibrators of known antigen values, 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 LI4014F1).

Bring reagents to room temperature before use.