Anti-SBCAD/ACADSB Antibody

• Catalog Number: orb669153
• Category: Antibodies
• Description: Anti-SBCAD/ACADSB Antibody. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Monkey, Mouse, Rat.
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, WB
• Reactivity: Human, Monkey, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human SBCAD/ACADSB, identical to the related mouse and rat sequences.
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 40-44 kDa
• UniProt ID: P45954
• 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.
• Alternative names: Annexin A8; Annexin VIII; Annexin-8; Vascular anti
• Note: For research use only
• Application notes: Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat, Monkey Immunocytochemistry/Immunofluorescence, 5μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human, Mouse, Rat. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of RAW264.7 cells using anti-SBCAD/ACADSB antibody. Overlay histogram showing RAW264.7 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-SBCAD/ACADSB 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.

Flow Cytometry analysis of RH-35 cells using anti-SBCAD/ACADSB antibody. Overlay histogram showing RH-35 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-SBCAD/ACADSB 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.

Flow Cytometry analysis of THP-1 cells using anti-SBCAD/ACADSB 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-SBCAD/ACADSB 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 SBCAD/ACADSB using anti-SBCAD/ACADSB antibody. SBCAD/ACADSB was detected in immunocytochemical section of Mcf-7 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-SBCAD/ACADSB Antibody overnight at 4°C. DyLight®488 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 SBCAD/ACADSB using anti-SBCAD/ACADSB 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 A431 whole cell lysates, Lane 2: human T47D whole cell lysates, Lane 3: human A549 whole cell lysates, Lane 4: monkey Cos-7 whole cell lysates, Lane 5: rat liver tissue lysates, Lane 6: mouse liver tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150 mA 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-SBCAD/ACADSB 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 1:5000 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 SBCAD/ACADSB at approximately 40-44 KD. The expected band size for SBCAD/ACADSB is at 47 KD.