Cytochrome P450 2c19/CYP2C19 (CYP2C19) Antibody – Mouse Monoclonal

Figure 1. Western blot analysis of Cytochrome p450 2C19/CYP2C19 using anti-Cytochrome p450 2C19/CYP2C19 antibody (MBS1754005).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: monkey liver tissue lysates
Lane 2: rat liver tissue lysates
Lane 3: mouse liver tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with mouse anti-Cytochrome p450 2C19/CYP2C19 antigen affinity purified monoclonal antibody (Catalog # MBS1754005) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for Cytochrome p450 2C19/CYP2C19 at approximately 55KD. The expected band size for Cytochrome p450 2C19/CYP2C19 is at 55KD.

Figure 2. IHC analysis of Cytochrome p450 2C19/CYP2C19 using anti-Cytochrome p450 2C19/CYP2C19 antibody (MBS1754005).
Cytochrome p450 2C19/CYP2C19 was detected in paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cytochrome p450 2C19/CYP2C19 Antibody (MBS1754005) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen.

Figure 3. Flow Cytometry analysis of A431 cells using anti- Cytochrome p450 2C19/CYP2C19 antibody (MBS1754005).
Overlay histogram showing A431 cells stained with MBS1754005 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cytochrome p450 2C19/CYP2C19 Antibody (MBS1754005, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Figure 4. Flow Cytometry analysis of U20S cells using anti- Cytochrome p450 2C19/CYP2C19 antibody (MBS1754005). Overlay histogram showing U20S cells stained with MBS1754005 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Cytochrome p450 2C19/CYP2C19 Antibody (MBS1754005, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.