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.
Conjugation:
Unconjugated
Alternative Names:
Diablo homolog, mitochondrial, IAP-binding protein with low pI, Second mitochondria-derived activator of caspase, Smac, DIABLO, SMAC
Smac/Diablo Rabbit Polyclonal Antibody
Clonality:
Polyclonal
Concentration:
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Form:
Lyophilized
Target:
Diablo IAP-binding mitochondrial protein
Application Dilute:
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/1x10 6 cells, Human
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 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. 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.
WB analysis of Smac/Diablo using anti-Smac/Diablo antibody.La
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 4C. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. 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 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. 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 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. 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 4C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37C. 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 4C, 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 develop
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