LS-C355232

antibody from LSBio

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

LS-C355232

Provider

LSBio

Product name

A2M / Alpha-2-Macroglobulin Antibody (clone F1-P1C11 #3) LS-C355232

Antibody type

Monoclonal

Description

Ion exchange chromatography

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

F1-P1C11 #3

Storage

Store at -20°C. Avoid freeze-thaw cycles.

Western blot

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

Western blot analysis of Alpha2-Macroglobulin antibody was performed by loading 5ul of Human Plasma and 5ul PageRuler Plus Prestained Protein Ladder onto a 4-20% Tris-Glycine polyacrylamide gel. 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. Alpha 2 Macroglobulin was detected at ~180kD using an Alpha2-Macroglobulin monoclonal antibody 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 at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico.

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

Western blot analysis of Alpha2-Macroglobulin antibody was performed by loading 1000ng, 500ng, 250ng and 125ng of a Human Alpha2-Macroglobulin Recombinant protein and 5ul PageRuler Plus Prestained Protein Ladder onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter and blocked with 5% BSA in TBST for at least 1 hour at room temperature. Alpha 2 Macroglobulin was detected at ~180kD using an Alpha2-Macroglobulin monoclonal antibody 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 at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura.

ELISA

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

Direct ELISA analysis of Alpha2-Macroglobulin was performed by coating wells of a 96-well plate with 100ul per well of Alpha2-Macroglobulin recombinant protein diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4C. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 100ul per well of a mouse Alpha2-Macroglobulin monoclonal antibody 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 100ul per well of an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:10,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate 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.