Smac/Diablo Antibody

SKU: orb334593

Description

Smac/Diablo Antibody

Images & Validation

• Tested Applications: ELISA, FC, ICC, IF, IHC, IP, WB
• Reactivity: Human, Mouse, Rat
• Application Notes:
  Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human Immunocytochemistry/Immunofluorescence, 5μg/ml, Human Immunoprecipitation, 0.5-2 μg/ml, Human Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human ELISA, 0.1-0.5μg/ml, -. 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: E. coli-derived human Smac/Diablo recombinant protein (Position: A56-D239). Human Smac/Diablo shares 88.6% amino acid (aa) sequence identity with mouse Smac/Diablo.
• Molecular Weight: 20 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

Diablo homolog, mitochondrial; IAP-binding protein with low pI; Second mitochondria-derived activator of caspase; Smac; DIABLO; SMAC

Flow Cytometry analysis of K562 cells using anti-Smac/Diablo antibody. Overlay histogram showing K562 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-Smac/Diablo 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 Smac/Diablo using anti-Smac/Diablo antibody. Smac/Diablo 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 2 µg/mL rabbit anti-Smac/Diablo Antibody overnight at 4°C. Cy3 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.

IHC analysis of Smac/Diablo using anti-Smac/Diablo antibody. Smac/Diablo was detected in a paraffin-embedded section of human testis 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 1 µg/ml rabbit anti-Smac/Diablo Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of Smac/Diablo using anti-Smac/Diablo antibody. Smac/Diablo was detected in a paraffin-embedded section of mouse testis 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 1 µg/ml rabbit anti-Smac/Diablo Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of Smac/Diablo using anti-Smac/Diablo antibody. Smac/Diablo was detected in a paraffin-embedded section of rat testis 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 1 µg/ml rabbit anti-Smac/Diablo Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of Smac/Diablo using anti-Smac/Diablo 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 30 ug of sample under reducing conditions. Lane 1: rat testis tissue lysates, Lane 2: 22RV1 whole cell lysates, Lane 3: SKOV whole cell 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-Smac/Diablo 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 Smac/Diablo at approximately 25 kDa. The expected band size for Smac/Diablo is at 27 kDa.

Quick Database Links

UniProt

Q9NR28