IL-1 beta Recombinant Mouse Monoclonal Antibody [A7F7-R]

☑ Cell treatment (CT)

Western blot analysis of IL-1 beta on different lysates with Mouse anti-IL-1 beta antibody (HA601036) at 1/1,000 dilution.

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

Lysates/proteins at 10 µg/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 (HA601036) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Western blot analysis of IL-1 beta on different proteins with Mouse anti-IL-1 beta antibody (HA601036) at 1/2,000 dilution.

Lane 1: Recombinant mouse IL-1 beta
Lane 2: Recombinant rat IL-1 beta
Lane 3: Recombinant human IL-1 beta

Lysates/proteins at 50 ng/Lane.

Exposure time: 30 seconds;

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 (HA601036) 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 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-IL-1 beta antibody (HA601036) 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 (HA601036) 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.

IL-1 beta Antibody (HA601036) 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 StartingBlock 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.

Application: IF-Tissue

Species: Human

Site: tonsil

Sample: Paraffin-embedded section

Antibody concentration: 1/200