PKC alpha Antibody (YA131)

Western blot analysis of extracts from HEK293 (lane 1(20μg)) 、Jurkat (lane 2(20μg)) 、NIH-3T3 (lane 3(20μg)) and C6 (lane 4(20μg)) using PKC alpha Antibody (HY-P80430) . 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 (Hsp90, 1/10000) 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 of extracts from HEK293 (lane 1(20μg)) 、HeLa (lane 2(20μg)) 、NIH-3T3 (lane 3(20μg)) and C6 (lane 4(20μg)) using PKC alpha Antibody (HY-P80430) . Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 1.5 hour at room temperature. The primary antibody (1/2000) and Loading control antibody (β Tubulin, 1/10000) 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 of extracts from WT HT-1080 Cells (lane 1(20μg)) and KD HT-1080 Cells (lane 2(20μg)) using PKC alpha Antibody (HY-P80430) . Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 1.5 hour at room temperature. The primary antibody (1/2000) and Loading control antibody (Vinculin, 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 of extracts from HEK293T(lane 2(20μg) , U87 (lane 3(20μg) ,NIH/3T3(lane 4(20μg)and C6( lane 5(20μg) using PKC alpha Antibody (HY-P80430). Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody ( 1/1000) and Loading control antibody (Beta Actin, HY-P80993，1/10000) 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 Colon cancer‌ tissue using PKC alpha 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-P80430, 1:100 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 Colon cancer‌ tissue using PKC alpha 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-P80430, 1:100 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 Endometrial Carcinoma tissue using PKC alpha 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-P80430, 1:100 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 Ovarian Cancer‌ tissue using PKC alpha 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-P80430, 1:100 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 Tonsil tissue using PKC alpha 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-P80430, 1:100 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 Liver Cancer tissue using PKC alpha 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-P80430, 1:100 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 Mouse brain tissue using PKC alpha Antibody (HY-P80430 , 1/200) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. 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.

Immunohistochemical analysis of paraffin-embedded Mouse Hippocampus tissue using PKC alpha Antibody (HY-P80430, 1/200) .The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. 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.

Immunohistochemical analysis of paraffin-embedded Human brain tissue using PKC alpha Antibody (HY-P80430, 1/200) .The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. 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.

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue using PKC alpha Antibody (HY-P80430, 1/200) .The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. 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.

Immunohistochemical analysis of paraffin-embedded Human spleen tissue using PKC alpha Antibody (HY-P80430, 1/200) .The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. 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.

Immunohistochemical analysis of paraffin-embedded Human colon tissue using PKC alpha Antibody (HY-P80430, 1/200) .The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (PH 6.0)for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. 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.

Flow cytometric analysis of 1X10⁶ HeLa cells labeling PKC alpha Antibody (HY-P80430, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/50 dilution for an hour at 4℃. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8002) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Rabbit IgG Isotype Control (HY-P80879, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black).

Immunocytochemistry analysis of HeLa cells labeling PKC alpha with PKC alpha Antibody (HY-P80430) at 1/50 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 PKC alpha Antibody (HY-P80430) at 1/50 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 PKC alpha with PKC alpha Antibody (HY-P80430) 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 PKC alpha Antibody (HY-P80430) 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).