Western blot analysis of hCG was performed by loading the indicated amounts of purified hCG protein, and 15ul of PageRuler Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane (Product # 88518) using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk for at least 1 hour at room temperature. hCG was detected at ~32 and 22 kD using a anti hCG antibody (Product # MA5-14701) at a concentration of 4ug/ml in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG (Fc) secondary antibody (Product # 31439) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Human Chorionic Gonadotropin|
|Storage buffer||0.01M potassium phosphate with 2.5mM KCl, 0.135M NaCl, 1mg/ml BSA|
|Contains||0.05% 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 *|
|ELISA (ELISA)||Assay Dependent|
|Radioimmune Assays (RIA)||Assay Dependent|
|Western Blot (WB)||1:250-1:500|
* 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.
Product MA514701 is a smaller package size of MIH9813 (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.