GM-CSF Mouse Monoclonal Antibody [A6C5]

Western blot analysis of GM-CSF on recombinant protein with Mouse anti-GM-CSF antibody (HA601033) at 1/2,000 dilution.

Lysates/proteins at 50 ng/Lane.

Predicted band size: 25 kDa
Observed band size: 25 kDa

Exposure time: 2 minutes;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601033) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

GM-CSF Antibody (HA601033) in indirect ELISA.

Indirect ELISA analysis of GM-CSF was performed by coating wells of a 96-well plate with 50 µl per well of GM-CSF antigen diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 50 µl per well of a mouse GM-CSF monoclonal antibody starting at a concentration of 20 µg/mL and serially diluting it to a concentration of 1.28 ng/mL for 2 hours at room temperature. The plate was washed and incubated with 50 µl 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 5 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.