CHD3 Antibody (YA945)

Western blot analysis was performed on extracts from Hela (lane 1, 15 μg), HepG2 (lane 2, 15 μg), and Jurkat (lane 3, 15 μg) using CHD3 Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non - fat milk in TBST at 4°C overnight. The primary antibody (1:1000 dilution) and the loading control antibody (beta-Tubulin(HRP), HY-P80955A, 1:5000 dilution) was incubated in 5% non-fat milk in TBST for 1 hour at 37°C. Goat Anti - Rabbit IgG - HRP Secondary Antibody (1:20000 dilution) was then applied for 40 minutes at 37°C.

Western blot analysis of extracts from K562 (lane 2(20μg), SiHa (lane 3(20μg), Jurkat (lane 4(20μg), NIH/3T3 (lane 5(20μg) using CHD3 Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA 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.

Flow Cytometry analysis of Hela cells labelling CHD3 (red) with CHD3 Antibody (anti-CHD3) (HY-P81264A). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then cells were stained with the primary antibody at 1/200 dilution for an hour at 4℃. Goat Anti-Rabbit IgG H&L (AF488) (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 CHD3 with CHD3 Antibody (HY-P81264A) 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 BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with CHD3 Antibody (HY-P81264A) at 1/50 dilution in BSA for Immunol Staining at 4 ℃ overnight. AF488-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).

Immunocytochemistry analysis of C6 cells labeling CHD3 with CHD3 Antibody (HY-P81264A) 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 BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with CHD3 Antibody (HY-P81264A) at 1/50 dilution in BSA for Immunol Staining at 4 ℃ overnight. AF488-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).