PARP1 Antibody (YA246)

Western blot analysis of extracts from Hela (lane 2(20μg) and Hela (lane 3(40μg)using PARP1 (HY-P80263) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% defatted milk powder in TBST for 2 hour at room temperature. The primary antibody (HY-P80263, 1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% defatted milk powder in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature.

Western blot analysis of extracts from HeLa (lane 1(20μg)) 、HepG2 (lane 2(20μg)) 、A549 (lane 3(20μg)) 、THP-1 (lane 4(20μg)) 、HEK293 (lane 5(20μg)) 、Jurkat (lane 6(20μg)) 、Raji (lane 7(20μg)) and Ramos (lane 8(20μg)) using PARP1 Antibody (HY-P80263) . 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 (1/1000) and Loading control antibody (GAPDH, 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.

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue using PARP1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80263, 1/4000) in 4℃ overnight. 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 colon tissue using PARP1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80263, 1/4000) in 4℃ overnight. 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 1X10^6 Hela cells labeling PARP1 Antibody (HY-P80263, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/1000 dilution for an hour at 4℃. Alexa Fluor® 488-conjugated AffiniPure 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 PARP1with PARP1antibody (HY-P80263) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with PARP1 antibody (HY-P80263) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure 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).