MIA1201

antibody from Invitrogen Antibodies

Targeting: A2M CPAMD5, FWP007, S863-7

Western blot (Supportive data in Antibodypedia)

ELISA (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Radioimmunoassay (Recommended by provider)

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. The MIA1201 immunogen is purified human plasma AMG. MIA1201 detects Alpha2-Macroglobulin which has a predicted molecular weight of approximately 162 kDa. MIA1201 was formerly sold as a Seradyn product.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

F1-P1C11 #3

Vial size

1 mg

Concentration

1 mg/mL

Storage

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

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.