Direct ELISA analysis of hCG proteins was performed by coating wells of a plate with a purified hCG protein extracted from human urine at a concentration of 10ug/ml overnight at 4C. The plate was washed 3 times with ELISA Wash Buffer (Product # N503) and blocked with 300 ul of StartingBlock (PBS) Blocking Buffer (Product # 37538) for at least 1 hour at room temperature. After washing 100ul of hCG monoclonal antibody (Product # MIH9827) was added to wells in duplicate at 10, 5, 2.5, 1.25, 0.62, 0.31, 0.15, 9.5, 0.078, 0.039, 0.019 and 0 ug/ml concentrations, and the samples were incubated for 2 hours at room temperature. The plate was washed, and then incubated with 100ul per well of an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:10,000 for 1 hour at room temperature, and washed again with ELISA Wash Buffer. The plate was developed by incubating 100ul per well of 1-Step Ultra TMB substrate (Product # 34028) per well for 10 minutes at room temperature in the dark. The reaction was stopped with 100ul per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550nm.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Chorionic Gonadotropin, b-Subunit|
|Storage buffer||PBS, pH 7.4|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay Dependent|
|Radioimmune Assays (RIA)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
The observed molecular weight of alpha-hCG and beta-hCG are approximately 20-22 kD and 27-32 kD respectively.
The molecular weight is higher than the predicted molecular weight due to post translational modification.
MIH9827 was formerly sold as a Seradyn product.
Human chorionic gonadotropin (hCG) is a placental hormone that stimulates secretion of the pregnancy-sustaining steroid progesterone. hCG is a member of a family of glycoprotein hormones includes pituitary hormones lutropin (luteinizing hormone;LH), follitropin (follicle-stimulating hormone; FSH) and thyrotropin (thyroid-stimulating hormone; TSH). Members of this family are all composed of disulfide-rich heterodimers with two dissimilar subunits namely alpha- and beta-subunits. Alpha subunit, which is common to all the members of the family within a given species, with distinct but homologous beta subunits which confer hormone specificities.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.