Cytochrome P450 3A4 Antibody (YA3715)

Western blot analysis of extracts from Rat liver(lane 1(40μg) ) using CYP3A4 antibody(30346P). Proteins were transferred to a NC membrane and blocked with 5% Skim milk in TBST for 2 hour at room temperature. The primary antibody ( 1/1000) and Loading control antibody (GAPDH，20035, 1/1000) was used in 5% Skim milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

Flow cytometric analysis of 1X10^6 HepG2 cells labeling CYP3A4 Antibody(30346P,green). Cells were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Then stained with the primary antibody at 1/400 dilution overnight at 4℃.Alexa Fluor® 488 Goat Anti-mouse IgG H&L (invitrogen A11001) was used as the secondary antibody at 1/1,000 dilution for 45 minutes at room temperature. Mouse IgG Isotype Control (Invitrogen 14-4714-B2, gray) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (red).

Immunocytochemistry analysis of HepG2 cells labeling CYP3A4 with CYP3A4 Antibody (HY-P84018) 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 CYP3A4 Antibody (HY-P84018) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Mouse IgG H&L（HY-P8005, 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 HepG2 cells labeling CYP3A4 with CYP3A4 Antibody (HY-P84018) at 1/100 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 CYP3A4 Antibody (HY-P84018)at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Mouse IgG H&L（HY-P8005,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).

Immunofluorescence analysis of Hela cells labeling CYP3A4 antibody (30346P) 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 1% BSA in 10% negative goat serum for 1 hour at room temperature.Cells were then incubated with CYP3A4 antibody (30346P) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Alexa Fluor® 488 Goat Anti-mouse IgG H&L (invitrogen A11001, green) was used as the secondary antibody at 1/500 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue)