Cytochrome P450 2B6/CYP2B6 Antibody

SKU: orb546299

Description

Anti-Cytochrome P450 2B6/CYP2B6 Antibody. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human.

Images & Validation

• Tested Applications: FC, ICC, IF, WB
• Reactivity: Human
• Application Notes:
  Western blot, 0.1-0.5μg/ml Immunocytochemistry/Immunofluorescence, 5μg/ml Flow Cytometry (Fixed), 1-3μg/1x106 cells. Add 0.2ml of distilled water will yield a concentration of 500ug/ml

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the N-terminus of human Cytochrome P450 2B6/CYP2B6.
• Molecular Weight: 56 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

Storage & Handling

• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

Cytochrome P450 2B6; 1, 4-cineole 2-exo-monooxygenase; CYPIIB6; Cytochrome P450 IIB1; CYP2B6

Flow Cytometry analysis of THP-1 cells using anti-Cytochrome P450 2B6/CYP2B6 antibody. Overlay histogram showing THP-1 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Cytochrome P450 2B6/CYP2B6 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of Cytochrome P450 2B6/CYP2B6 using anti-Cytochrome P450 2B6/CYP2B6 antibody. Cytochrome P450 2B6/CYP2B6 was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL rabbit anti-Cytochrome P450 2B6/CYP2B6 Antibody overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of CYP2B6 using anti-CYP2B6 antibody. 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 50 ug of sample under reducing conditions. Lane 1: human Caco-2 whole cell lysates, Lane 2: human HEK293 whole cell lysates, Lane 3: human PC-3 whole cell lysates, Lane 4: human HL-60 whole cell lysates, Lane 5: human K562 whole cell lysates, Lane 6: human A549 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-CYP2B6 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CYP2B6 at approximately 56KD. The expected band size for CYP2B6 is at 56KD.

Quick Database Links

UniProt

P20813