MIA1201

antibody from Invitrogen Antibodies

Targeting: A2M CPAMD5, FWP007, S863-7

Western blot (Supportive data in Antibodypedia)

ELISA (Supportive data in Antibodypedia)

Antibody Data

Product number

MIA1201

Provider

Invitrogen Antibodies

Product name

A2M Monoclonal Antibody (F1-P1C11 #3)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MIA1201 targets Alpha2-Macroglobulin in ELISA, immunoprecipitation, and radioimmunoassay applications and shows reactivity with Human samples.

Antibody clone number

F1-P1C11 #3

Concentration

1 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Alpha2-Macroglobulin antibody was performed by loading 1000 ng, 500 ng, 250 ng and 125 ng of a Human Alpha2-Macroglobulin Recombinant protein and 5 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% BSA in TBST for at least 1 hour at room temperature. Alpha 2 Macroglobulin was detected at ~180 kD using an Alpha2-Macroglobulin monoclonal antibody (Product # MIA1201 F1-P1C11 #3) at a concentration of 2 µg/mL in 5% BSA in TBST overnight at 4C on a rocking platform, followed by an incubation with a goat anti-mouse IgG-HRP Superclonal secondary antibody (Product # A28177) at a dilution of 1:1000 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 Alpha2-Macroglobulin antibody was performed by loading 5 µL of Human Plasma and 5 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% milk in TBST for at least 1 hour at room temperature. Alpha 2 Macroglobulin was detected at ~180 kD using an Alpha2-Macroglobulin monoclonal antibody (Product # MIA1201 F1-P1C11 #3) at a concentration of 1 µg/mL in 5% milk in TBST overnight at 4C on a rocking platform, followed by an incubation with a goat anti-mouse IgG-HRP Superclonal secondary antibody (Product # A28177) at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).

ELISA

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Direct ELISA analysis of Alpha2-Macroglobulin was performed by coating wells of a 96-well plate with 100 µL per well of Alpha2- Macroglobulin recombinant protein diluted in carbonate/bicarbonate buffer (Product # 28382), at a concentration of 1 µg/mL overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer (Product # 37538), and incubated with 100 µL per well of a mouse Alpha2-Macroglobulin monoclonal antibody (Product # MIA1201 F1-P1C11 #3) starting at a concentration of 2 µg/mL and serially diluting it to a concentration of 0.03125 µg/mL for 2 hours at room temperature. The plate was washed and 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 one hour at room temperature. Detection was performed using an Ultra TMB Substrate (Product # 34028) for 30 minutes at room temperature in the dark. The reaction was stopped with 0.16M sulfuric acid and absorbances were read on a spectrophotometer at 450-550 nm.