Bcl-2 Antibody (YA589)

MedchemExpress Validation
Western blot analysis of extracts from HeLa (lane 1(20μg)) 、SH-SY5Y (lane 2(20μg)) 、HepG2 (lane 3(20μg)) 、HEK293 (lane 4(20μg)) 、Jurkat (lane 5(20μg)) 、HL-60 (lane 6(20μg)) 、U937 (lane 7(20μg)) and MCF7 (lane 8(20μg)) using Bcl-2 Antibody (HY-P80029) . 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 (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.

MedchemExpress Validation
Western blot analysis of extracts from HeLa (lane 1(20μg)) 、SH-SY5Y (lane 2(20μg)) 、HepG2 (lane 3(20μg)) 、HEK293 (lane 4(20μg)) 、Jurkat (lane 5(20μg)) 、HL-60 (lane 6(20μg)) 、U937 (lane 7(20μg)) and MCF7 (lane 8(20μg)) using Bcl-2 Antibody. 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 (HY-P80029, 1/1000) , competitor's antibody (1/1000) 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.

MedchemExpress Validation
Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue using Bcl-2 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) 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-P80029, 1/2000) 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.

MedChemExpress Validation
Flow cytometric analysis of 1X10⁶ Jurkat cells labeling Bcl-2 Antibody (HY-P80029, red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/500 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).

MedchemExpress Validation
Immunocytochemistry analysis of Hela cells labeling Bcl-2 with Bcl-2 Antibody (HY-P80029) 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 Bcl-2 Antibody (HY-P80029)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).

MedChemExpress Validation
Immunocytochemistry analysis of A549 cells labeling Bcl-2 with Bcl-2 Antibody (HY-P80029) 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 Bcl-2 Antibody (HY-P80029) 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).