- LS-C355235 - LSBio APOA1 antibody

No submitted references: Submit reference

No comments: Submit comment

Supportive validation

Supportive validation

Submitted by: LSBio (provider)
Main image
Experimental details: Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100ul of an Apo A-1 rabbit oligoclonal antibody diluted to a concentration of 1 µg/mL in carbonate/bicarbonate buffer overnight at 4C. Wells were blocked with 150ul of StartingBlock T20 (TBS) Blocking Buffer for 30 minutes, and 80ul of recombinant human Apo A-1 or recombinant mouse Apo A-1 was added to the plate at concentrations ranging from 1.6-1000ng/ml and incubated for 1 hour at room temperature. The plate was washed with 1X TBST, and 100ul per well of an Apo A-1 mouse monoclonal antibody was added to each well for 1 hour at room temperature. The plate was washed, and 100ul per well of an HRP-conjugated rabbit anti-mouse IgG cross-adsorbed secondary antibody was incubated for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate, followed by Stop Solution. Absorbances were read on a spectrophotometer at 450-550nm.

Supportive validation

Submitted by: LSBio (provider)
Main image
Experimental details: Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100ul of an Apo A-1 rabbit oligoclonal antibody at a concentration of 1 µg/mL in carbonate/bicarbonate buffer overnight at 4C. Wells were blocked with 150ul of StartingBlock T20 (TBS) Blocking Buffer for 30 minutes, and 80ul of recombinant human Apo A-1 standards ranging from 1.6-1000ng/ml (A) or 100ul of diluted normal human serum or diluted serum from a patient with dyslipidemia (B) were incubated for 1 hour at room temperature. The plate was washed with 1X TBST, and 100ul per well of an Apo A-1 mouse monoclonal antibody was added to each well for 1 hour at room temperature. The plate was washed, and 100ul per well of a biotinylated rabbit anti-mouse IgG Superclonal secondary antibody was incubated for 1 hour, followed by Streptavidin-HRP for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate, followed by Stop Solution. Absorbances were read on a spectrophotometer at 450-550nm.

Supportive validation

Submitted by: LSBio (provider)
Main image
Experimental details: Immunohistochemistry was performed on human liver tissue sections. To expose Apo A-1, tissues were microwaved for 10 minutes in 10mM sodium citrate, pH 6.0. Following antigen retrieval, endogenous peroxidases were blocked in a 3% hydrogen peroxide-methanol solution for 15 minutes, and then tissues were blocked in 3% BSA in PBS for 30 minutes at room temperature. Tissues were probed with an Apo A-1 monoclonal antibody. Proteins were transferred to a Nitrocellulose Membrane using the G2 Fast Blotter, and blocked with 5% milk in TBST for at least 1 hour at room temperature. Apo A-1 was detected at ~50kD using an Apolipoprotein A-1 monoclonal antibody at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG Fc-specific secondary antibody at a dilution of 1:40,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura.

LS-C355235