Technical Data
Beta-Chorionic Gonadotropin, Free, Human (hCGbeta) BioAssay™ ELISA Kit
Kits and Assays Storage: 4°CShipping: Blue Ice
Intended Use:
For the quantitative determination of free beta subunit of human chorionic gonadotropin (free Beta-hCG) concentration in human serum.

Standard Size: 96 wells

Range: 0-50ng/ml

Sensitivity: 2.5ng/ml

Crossreactivity: This kit has been validated for use with human serum samples only.

Principle of the Test:
The free beta-hCG ELISA test is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a unique monoclonal antibody directed against a distinct antigenic determinant on the free beta-hCG. Mouse monoclonal anti-free-beta-hCG antibody is used for solid phase immobilization (on the microtiter wells). A goat anti-whole hCG antibody is in the antibody-enzyme (horseradish peroxidase) conjugate solution. The test sample is allowed to react sequentially with the two antibodies, resulting in the free beta-hCG molecules being sandwiched between the solid phase and enzyme-linked antibodies. After two separate 30 minute incubations at 37°C, the wells are washed with water to remove unbound labeled antibodies. A solution of TMB Reagent is added and incubated for 20 minutes, resulting in the development of a blue color. The color development is stopped with the addition of Stop Solution changing the color to yellow. The concentration of beta-hCG is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450nm.

Kit Components:
C5069-68A: Microtiter Plate, 1x96wells Coated with murine monoclonal anti-free beta-hCG antibody
C5069-68B: Calibrator 0ng/ml, 1x1vial
C5069-68C: Calibrator 2.5ng/ml, 1x1vial
C5069-68D: Calibrator 5ng/ml, 1x1vial
C5069-68E: Calibrator 10ng/ml, 1x1vial
C5069-68F: Calibrator 25ng/ml, 1x1vial C5069-68G: Calibrator 50ng/ml, 1x1vial C5069-68H: Sample Diluent, 1x13ml
C5069-68J: Pab (HRP), 1x18ml
C5069-68K: TMB Solution, 1x11ml
C5069-68L: Stop Solution, 1N HCl, 1x11ml

Storage and Stability:
Store kit components at 4°C. Unopened kit components are stable for 6 months at 4°C. Reconstituted standards are stable for up to 30 days when stored sealed at 4°C.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
1. Engall, E., Enzyme immunoassay ELISA and EMIT. In: Van Vunakis, H. and Langone, J.J. (eds.), Methods in Enzymology, Academic Press, New York, 1980; 70:419-439. 2. Densem, J., and Wald, N.J., The stability of blood samples for the measurement of the free beta subunit of chorionic gonadotrophin. Prenat. Diagn., 1995; 15: 94-95. 3. Hay, D.L., Placental histology and the production of human choriogonadotrophin and its subunits in pregnancy. Br. J Obstet. Gynaecol., 1988; 95:1268-1275. 4. Ozturk, M., Bellet, D., Manil, L., et. al., Physiological studies of human chorionic gonadotropin (hCG), alpha hCG, and beta hCG as measured by specific monoclonal immunoradiometric assays. Endocrinology, 1987; 120:549-558. 5. Ozturk, M., Berkowitz, R., Goldstein, D., Bellet, D., Wands, J. R., Differential production of human chorionic gonadotropin and free subunits in gestational trophoblastic disease. Am. J. Obstet. Gynecol.,1988; 158:193-198. 6. Cole, L.A., Hartle, R.J., Laferla, J.J., et al., Detection of the free beta subunit of human chorionic gonadotropin (hCG) in cultures of normal and malignant trophoblast cells, pregnancy sera, and sera of patients with choriocarcinoma. Endocrinology, 1983; 113:1176-1178. 7. Gaspard, U.J., Reuter, A.M., Devėlle, J-L, et al., Serum concentration of human chorionic gonadotropin and its alpha and beta subunits. Clin. Endocrinol. (OXF), 1980; 13:319-329.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.