MIH9812

antibody from Invitrogen Antibodies

Targeting: CGB8

ELISA

Immunoprecipitation

Radioimmunoassay

Antibody data

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Validation data

Product number: MIH9812 - Provider product page
Provider: Invitrogen Antibodies
Product name: hCG Monoclonal Antibody (SC1)
Antibody type: Monoclonal
Antigen: Other
Description: The observed molecular weight of alpha-hCG and beta-hCG are approximately 20-22 kD & 27-32 kD respectively. The molecular weight is higher than the predicted molecular weight due to post translational modification. MIH9812 was formerly sold as a Seradyn product.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: SC1
Vial size: 1 mg
Concentration: 5 mg/mL
Storage: Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C

CGB8 protein structure - MIH9812 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of hCG was performed by loading the indicated amounts of protein purified hCG protein extracted from human urine, and 15 µL 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 & 22 kD using a anti hCG antibody (Product # MIH9812) at a concentration of 4 µg/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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of hCG was performed by loading the indicated amounts of protein purified hCG protein extracted from human urine, and 15 µL 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 & 22 kD using a anti hCG antibody (Product # MIH9812) at a concentration of 4 µg/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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of hCG proteins was performed by spiking 5 ugs of purified hCG protein extracted from human urine into 1 mL of 1:2 diluted human urine sample. Antigen-antibody complexes were formed by incubating with 5 µg of hCG antibody (Product # MIH9812) overnight on end over end tube rotator at 4C. The immune complexes were captured by using 50 µL Protein A/G Agarose (Product # 20421), washed extensively, and eluted with Lane Marker Reducing Sample Buffer (Product # 39000). Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane (Product # 88518), and blocked with 5% milk for at least 1 hour at room temperature. hCG was detected at ~32 & 22 kD using hCG monoclonal antibody (Product # MA5-14680) at a concentration of 4 µg/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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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 10 µg/mL overnight at 4C. The plate was washed 3 times with ELISA Wash Buffer (Product # N503) and blocked with 300 µL of StartingBlock (PBS) Blocking Buffer (Product # 37538) for at least 1 hour at room temperature. After washing 100 µL of hCG monoclonal antibody (Product # MIH9812) 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 µg/mL concentrations, and the samples were incubated for 2 hours at room temperature. The plate was washed, and then incubated with 100 µL 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 100 µL 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 100 µL per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.