IL-1 beta/IL-1F2 Antibody (A7F7-R)

ELISA: IL-1 beta/IL-1F2 Antibody (A7F7-R) [NBP3-32464]

ELISA: IL-1 beta/IL-1F2 Antibody (A7F7-R) [NBP3-32464] - IL-1 beta/IL-1F2 Antibody (NBP3-32464) in indirect ELISA.

Indirect ELISA analysis of IL-1 beta was performed by coating wells of a 96-well plate with 50 µl per well of IL-1 beta antigen diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with blocking buffer, and incubated with 50 µl per well of a mouse IL-1 beta 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.

Western Blot: IL-1 beta/IL-1F2 Antibody (A7F7-R) [NBP3-32464]

Western Blot: IL-1 beta/IL-1F2 Antibody (A7F7-R) [NBP3-32464] - Western blot analysis of IL-1 beta/IL-1F2 on different lysates with Mouse anti-IL-1 beta/IL-1F2 antibody (NBP3-32464) at 1/1,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: THP-1 treated with 80nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300ng/mL BFA for 3 hours cell lysate

Lysates/proteins at 10 ug/Lane.

Predicted band size: 31 kDa
Observed band size: 31 kDa

Exposure time: 1 minute;

4-20% 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 (NBP3-32464) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: IL-1 beta/IL-1F2 Antibody (A7F7-R) [NBP3-32464]

Immunohistochemistry: IL-1 beta/IL-1F2 Antibody (A7F7-R) [NBP3-32464] - Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-IL-1 beta/IL-1F2 antibody (NBP3-32464) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32464) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.