Human IL-2 Mouse Monoclonal Antibody [A8B7] - BSA and Azide free (Detector)

Human IL-2 Antibody (HA601006) in indirect ELISA.

Indirect ELISA analysis of Human IL-2 was performed by coating wells of a 96-well plate with 50 µl per well of Human IL-2 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 Human IL-2 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.

Standard curve of human IL-2 matched pair antibodies:

Sandwich ELISA analysis of human IL-2 matched pair antibodies
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody [PSH01-42] diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1%BSA blocking buffer, and incubated with serial diluted IL-2 protein starting from 2500 pg/ml to 0 pg/ml and detect antibody [A8B7]-Biotin (0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Western blot analysis of Human IL-2 on IL-2 recombinant protein with Mouse anti-Human IL-2 antibody (HA601006) at 1/500 dilution.

Lysates/proteins at 50 ng/Lane.

Exposure time: 30 seconds;

15% 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 (HA601006) at 1/500 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.