CYP7A1 Antibody – Rabbit Polyclonal

Figure 1. Western blot analysis of CYP7A1 using anti-CYP7A1 antibody (MBS1753447).
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: rat liver tissue lysates
Lane 2: mouse liver tissue lysates
Lane 3: monkey liver tissue lysates
Lane 4: human HepG2 whole cell lysates
Lane 5: human Hele whole cell 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 rabbit anti-CYP7A1 antigen affinity purified polyclonal antibody (Catalog # MBS1753447) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for CYP7A1 at approximately 54KD. The expected band size for CYP7A1 is at 54KD.

Figure 2. Flow Cytometry analysis of U87 cells using anti-CYP7A1 antibody (MBS1753447).
Overlay histogram showing U87 cells stained with MBS1753447 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CYP7A1 Antibody (MBS1753447, 1μg/1x10⁶ cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.