LH ELISA Assay Kit

$230.00

The Eagle Biosciences LH ELISA Assay Kit is a solid phase enzyme immunoassay for the quantitative determination of the luteinizing hormone (LH) concentration in human serum or plasma. The LH ELISA Assay Kit is intended for research use only and not for diagnostic procedures.

SKU: DCM009 Categories: , ,

LH ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.22 mIU/ml
Dynamic Range: 5 – 200 mIU/ml
Incubation Time: 1.5 hour
Sample Type: Serum, Plasma
Sample Size: 20 µL

Controls Included

Product Developed and Manufactured in Italy by Diametra

Additional Information

Assay Background

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 β-subunit is unique. The β-subunit confers the biological activity to the molecule.

The α-subunit consists of 89 amino acid residues while the β-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.

Assay Principle

In this LH ELISA Assay Kit, the LH calibrators, the patient specimens and/or the controls (containing the native antigen) are first added to streptavidin coated wells. Then monoclonal biotinylated 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. A reaction between the various LH antibodies and native LH occurs in the microwells without competition or steric hindrance forming a soluble sandwich complex. The interaction is illustrated in the following equation:

ka

EnzAb + AgLH + BtnAb(m) « EnzAb – AgLH-BtnAb(m)

     k-a

BtnAb(m) = biotinylated monoclonal antibody (excess quantity)
AgLH = native antigen (variable quantity)
EnzAb = enzyme labeled antibody (excess quantity)
EnzAb-AgLH-BtnAb(m) = antigen-antibodies sandwich complex
>ka = rate constant of association
k-a = rate constant of dissociation

Simultaneously, the complex is deposited to the well through the high affinity reaction of streptavidin and biotinylated antibody. This interaction is illustrated below:

EnzAb -AgLH-BtnAb(m) + StreptavidinC.W. Immobilized complex

Streptavidin C.W. = streptavidin immobolized on well.

Immobilized complex =antibodies-antigen sandwich bound.

After equilibrium is attained, the antibody-bound fraction is separated from unbound antigen by a washing step. 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 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.

Manual

Product Manual