MA5-14668

antibody from Invitrogen Antibodies

Targeting: APOA1

Western blot (Data presented on provider website)

ELISA (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Radioimmunoassay (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-14668

Provider

Invitrogen Antibodies

Product name

ApoA1 Monoclonal Antibody (311)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

By sandwich ELISA, MA5-14668 can be used as a detection antibody with Product # 710263 or # 701239 as a coating antibody, to generate a matched pair. Using these matched pairs, recombinant human Apo A-1, but not recombinant mouse Apo A-1, was detected. MA5-14668 can be used to detect Apo A-1 from serum samples. To increase sensitivity of sandwich ELISAs with MA5-14668, a biotinylated detection antibody followed by Streptavidin-HRP is recommended. By Western blot, MA5-14668 detects recombinant human Apo A-1, but not recombinant mouse Apo A-1. MA5-14668 may not be as successful for detecting endogenous Apo A-1 by Western blot. Product MA514668 is a smaller package size of MIA1402 (formerly sold as a Seradyn product).

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

311

Vial size

100 μg

Concentration

1 mg/mL

Storage

Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C

ELISA

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 rabbit monoclonal antibody (Product # 701239) diluted to a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) or recombinant mouse Apo A-1 (Product # 50918-M02H-50) was added to the plate at concentrations ranging from 1.6-1000 ng/mL and incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100 µL per well of an Apo A-1 mouse monoclonal antibody (Product # MA5-14668) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of an HRP-conjugated rabbit anti-mouse IgG cross-adsorbed secondary antibody (Product # 31452) was incubated for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 Recombinant Rabbit Polyclonal Antibody (Product # 710263) diluted to a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) or recombinant mouse Apo A-1 (Product # 50918-M02H-50) was added to the plate at concentrations ranging from 1.6-1000 ng/mL and incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100 µL per well of an Apo A-1 Mouse Monoclonal Antibody (Product # MA5-14668) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of an HRP-conjugated Rabbit anti-Mouse IgG cross-adsorbed secondary antibody (Product # 31452) was incubated for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 rabbit monoclonal antibody (Product # 701239) at a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) standards ranging from 1.6-1000 ng/mL (A) or 100 µL 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 (Product # 28360), and 100 µL per well of an Apo A-1 mouse monoclonal antibody (Product # MA5-14668) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of a biotinylated rabbit anti-mouse IgG Superclonal™ secondary antibody (Product # A27024) was incubated for 1 hour, followed by Streptavidin-HRP (Product # 21126) for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 Recombinant Rabbit Polyclonal Antibody (Product # 710263) at a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) standards ranging from 1.6-1000 ng/mL (A) or 100 µL 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 (Product # 28360), and 100 µL per well of an Apo A-1 Mouse Monoclonal Antibody (Product # MA5-14668) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of a biotinylated Rabbit anti-Mouse IgG Superclonal secondary antibody (Product # A27024) was incubated for 1 hour, followed by Streptavidin-HRP (Product # 21126) for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 Recombinant Rabbit Polyclonal Antibody (Product # 710263) diluted to a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) or recombinant mouse Apo A-1 (Product # 50918-M02H-50) was added to the plate at concentrations ranging from 1.6-1000 ng/mL and incubated for 1 hour at room temperature. The plate was washed with 1X TBST (Product # 28360), and 100 µL per well of an Apo A-1 Mouse Monoclonal Antibody (Product # MA5-14668) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of an HRP-conjugated Rabbit anti-Mouse IgG cross-adsorbed secondary antibody (Product # 31452) was incubated for 30 minutes at room temperature. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 Recombinant Rabbit Polyclonal Antibody (Product # 710263) at a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) standards ranging from 1.6-1000 ng/mL (A) or 100 µL 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 (Product # 28360), and 100 µL per well of an Apo A-1 Mouse Monoclonal Antibody (Product # MA5-14668) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of a biotinylated Rabbit anti-Mouse IgG Superclonal secondary antibody (Product # A27024) was incubated for 1 hour, followed by Streptavidin-HRP (Product # 21126) for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA of Apolipoprotein A-1 was performed by coating wells of a 96-well plate with 100 µL of an Apo A-1 rabbit monoclonal antibody (Product # 701239) at a concentration of 1 µg/mL in carbonate/bicarbonate buffer (Product # 28382) overnight at 4C. Wells were blocked with 150 µL of StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 30 minutes, and 80 µL of recombinant human Apo A-1 (Product # 10686-H02H-50) standards ranging from 1.6-1000 ng/mL (A) or 100 µL 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 (Product # 28360), and 100 µL per well of an Apo A-1 mouse monoclonal antibody (Product # MA5-14668) was added to each well for 1 hour at room temperature. The plate was washed, and 100 µL per well of a biotinylated rabbit anti-mouse IgG Superclonal? secondary antibody (Product # A27024) was incubated for 1 hour, followed by Streptavidin-HRP (Product # 21126) for 30 minutes. Detection was performed using 1-Step Ultra TMB Substrate (Product # 34028), followed by Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.