Prosurfactant Protein C Antibody (YA6950)

Western blot analysis of extracts from Human lung (lane 1(40μg)) 、Mouse lung (lane 2(40μg)) and Rat lung (lane 3(40μg)) using Prosurfactant Protein C Antibody (HY-P87267) . Proteins were transferred to a PVDF membrane and blocked with 5% nonfat dry milk in TBST for 1.5 hour at room temperature. The primary antibody (1/2000) and Loading control antibody (β Tubulin, 1/5000) was used in 5% nonfat dry milk in TBST at 4℃ overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

Western blot analysis was performed on protein extracts from Mouse lung (lane 2, 20 μg), Mouse lung (lane 3, 40 μg), Rat lung (lane 4, 20 μg), and Rat lung (lane 5, 40 μg) using Prosurfactant Protein C 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 (1:5000) and loading control antibody GAPDH Antibody (HRP) (HY-P80954A) (1:5000) 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 Lung Adenocarcinoma tissue using Prosurfactant Protein C antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P87267, 1:1000 dilution) at room temperature for 60 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 Lung Adenocarcinoma tissue using Prosurfactant Protein C antibody was performed. The section was pretreated using high-temperature and high-pressure mediated EDTA antigen retrieval buffer (pH 9.0), for 5 minutes. The tissues were incubated with primary antibody (HY-P87267, 1:1000 dilution) at room temperature for 60 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.

Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Lung Adenocarcinoma tissue using Prosurfactant Protein C antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P87267, 1:1000 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.

Tyramide signaling amplification based immunofluorescence was performed on paraffin-embedded human Lung Adenocarcinoma tissue using Prosurfactant Protein C antibody. Antigen retrieval was performed in EDTA buffer pH 9.0 (95 °C, 20 min) followed by cooling to RT. Then incubated with primary antibody (HY-P87267, 1:1000 dilution) at room temperature for 60 minutes and HRP conjugated secondary antibody for 10 minutes. Fluorescence was then developed with Vari Fluor 532 TSA (200×)（HY-D1832）. The tissues were counterstained with DAPI and mounted with Anti-fade mounting medium.