Phospho-PRC1 (Thr481) Antibody (YA1713)

Western blot analysis of extracts from HeLa(lane 2(20μg), Jurkat(lane 3(20μg), HUVEC(lane 4(20μg), HCT116(lane 5(20μg) using Phospho-PRC1 (Thr481) Antibody (HY-P81968). Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hours at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80993, 1/10,000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HY-P8001, 1/10,000) was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human Tonsil tissue using Phospho-PRC1 (Thr481) 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 overnight at 4°C with the primary antibody (HY-P81968, 1:50 dilution). 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 Tonsil tissue using Phospho-PRC1 (Thr481) 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 overnight at 4°C with the primary antibody (HY-P81968, 1:50 dilution). 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 Tonsil tissue using Phospho-PRC1 (Thr481) 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 overnight at 4°C with the primary antibody (HY-P81968, 1:50 dilution). 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 Testis tissue using Phospho-PRC1 (Thr481) 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 overnight at 4°C with the primary antibody (HY-P81968, 1:50 dilution). 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 Ovarian Cancer‌ tissue using Phospho-PRC1 (Thr481) 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 overnight at 4°C with the primary antibody (HY-P81968, 1:50 dilution). 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 Ovarian Cancer‌ tissue using Phospho-PRC1 (Thr481) 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 overnight at 4°C with the primary antibody (HY-P81968, 1:50 dilution). 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 Tonsil tissue using Phospho-PRC1 (Thr481) 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-P81968, 1:50 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 Tonsil tissue using Phospho-PRC1 (Thr481) 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-P81968, 1:50 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 Tonsil tissue using Phospho-PRC1 (Thr481) 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-P81968, 1:50 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 Testis tissue using Phospho-PRC1 (Thr481) 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-P81968, 1:50 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 Testis tissue using Phospho-PRC1 (Thr481) 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-P81968, 1:50 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 Testis tissue using Phospho-PRC1 (Thr481) 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-P81968, 1:50 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.

Immunocytochemistry analysis of HeLa cells labeling Phospho-PRC1 (Thr481) with Phospho-PRC1 (Thr481) Antibody (HY-P81968) at 1/100 dilution . Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Phospho-PRC1 (Thr481) Antibody (HY-P81968) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green） was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

Immunocytochemistry analysis of HeLa cells labeling Phospho-PRC1 (Thr481) with Phospho-PRC1 (Thr481) Antibody (HY-P81968) at 1/200 dilution . Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with Phospho-PRC1 (Thr481) Antibody (HY-P81968) at 1/200 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green） was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).