IL-1 Receptor Antagonist Protein Antibody (YA3095)

Western blot analysis was performed on extracts from A431 (lane 1, 15 μg), K562 (lane 2, 15 μg), Neuro-2a (lane 3, 15 μg), Jurkat (lane 4, 15 μg), Hela (lane 5, 15 μg), and 3T3 (lane 6, 15 μg) using IL-1RA Protein Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non - fat milk in TBST at 4°C overnight. The primary antibody (1:1000 dilution) and the loading control antibody (beta-Actin(HRP), HY-P80993, 1:5000 dilution) was incubated in 5% non-fat milk in TBST for 1 hour at 37°C. Goat Anti - Rabbit IgG - HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

Western blot analysis was performed on protein extracts (25 μg) from HepG2 (lane 2), HeLa (lane 3), HT-1080 (lane 4), K562 (lane 5), U-87 (lane 6), and MCF-7 (lane 7) using IL-1 Receptor Antagonist Protein antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot^® Turbo^™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. Thhe primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:2500) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005).

Immunohistochemical analysis of paraffin-embedded human mammary gland tissue using IL-1 Receptor Antagonist Protein antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P83350, 1:600 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

Immunohistochemical analysis of paraffin-embedded human appendix‌ tissue using IL-1 Receptor Antagonist Protein antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P83350, 1:600 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

Immunohistochemical analysis of paraffin-embedded human Liver tissue using IL-1 Receptor Antagonist Protein antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P83350, 1:600 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

Immunohistochemical analysis of paraffin-embedded human thyroid tissue using IL-1 Receptor Antagonist Protein antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P83350, 1:600 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

Immunohistochemical analysis of paraffin-embedded human Colorectal cancer tissue using IL-1 Receptor Antagonist Protein antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P83350, 1:600 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.

Immunohistochemical analysis of paraffin-embedded human placenta tissue using IL-1 Receptor Antagonist Protein antibody was performed. The section was pretreated using high-temperature mediated EDTA antigen retrieval buffer (pH 9.0), for 20 minutes. The tissues were incubated with primary antibody (HY-P83350, 1:600 dilution) at room temperature for 20 minutes. Detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. The tissues were counterstained with hematoxylin and mounted with neutral balsam mounting medium.