HCG Titration ELISA Kit

Beta HCG (Titration) ELISA kit description:

he Diagnostic Automation β-hCG EIA test kit is mainly intended to quantitatively determine Total beta-hCG concentration in human serum.

Materials provided with Beta HCG (Titration) ELISA Test Kit:

1. Anti-hCG Antibody-coated ELISA Microplate (96 wells)

2. enzyme conjugate reagent: 2 Vials contains anti-hCG-HRP with preservative buffer, ready to use.(1 Vials, 12 ml)

3. standard set, contains 0, 50, 250, 1000, 2500, 5000 & 10000 IU/L (6 vials, 1.5 ml & zero STD, 5 ml)

4. TMB Substrate: Hydrogen peroxide and tetra-methyl benzidine in preservative buffer, ready to use. (1 vial of 12ml)

5. Stop Solution: a normal hydrochloric acid (1 vial, 6 ml).

6. Concentrated washing buffer: PBS-Tween (1 vial 30ml) (20X)

7. Control serum: hCG in serum with preservatives(2Vial, 1.5 ml)

Materials Required, not Provided:

1. Precision pipettes(20,50,100,200 µl)

2. Distilled or deionized water

3. EIA kit Microplate Washer

4. EIA kit Microplate Reader with a 450 nm, If possible, 630 nm as reference.

5. Absorbent paper

Introduction

Human chorionic gonadotropin (hCG) with a molecular mass of 38,000 Da is a hormone synthesized by the placental trophoblast cells during pregnancy. It is heterodimeric, with an α (alpha) and β (beta) subunit. α subunit is identical to that of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and human HCG, and β subunit is responsible to specific hormone effects of hCG molecule. Due to constructional similarities, physiological and immunological characteristics, hCG is almost identical to LH hormone. hCG has leutropic effect and supports the formation of corpus luteum; also, hCG determines the hormonal function of testes and adrenal glands of fetus. Serum hCG appears in the early stages of pregnancy, so that its concentration increases and reaches to its peak in the first trimester and then progressively decreases until the child birth. Therefore, measurement of hCG is important for monitoring of pregnancy particularly in the first trimester. In addition to early detection of pregnancy, hCG measurement is important in diagnosis of abnormal pregnancy. hCG has a slow rise and remain at lower levels in an ectopic compared to normal pregnancy. High concentrations of hCG generally can be observed in multiple pregnancy and in patients with trophoblast and non-trophoblast tumors such as testicular, breast, bronchial carcinoma and embryonic cells.

Principle of the assay

In this ELISA test, kit is designed based on immunoenzymometric assay. This ELISA assay is based on use of sandwich method and enzyme immunological method. Anti-hCG monoclonal antibody is used to conjugation with HRP enzyme for coating between the solid phase and an Anti-hCG antibody. By addition of the sample serum, the HCG molecule reacts with mentioned antibodies and sandwiches between bounded antibody on the solid phase and conjugated antibody. After incubation period, the sample is then decanted and the wells are washed. Then an enzyme substrate is added to each well. Standards with specified concentrations are tested with unknown sample and the concentration of unknown samples is achieved Based on the standard curve of light absorption according to the concentration of hCG. The activity of the enzyme has a direct relationship with hCG in the samples.