Anti-SIAH Interacting Protein/CACYBP Antibody Picoband®

Figure 1. Western blot analysis of CACYBP using anti-CACYBP antibody (RP1104).
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: human MCF-7 whole cell lysates,
Lane 2: human RT4 whole cell lysates,
Lane 3: human SW620 whole cell lysates,
Lane 4: human U251 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat brain tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse brain 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-CACYBP antigen affinity purified polyclonal antibody (Catalog # RP1104) 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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CACYBP at approximately 26 kDa. The expected band size for CACYBP is at 26 kDa.

Figure 2. IHC analysis of CACYBP using anti-CACYBP antibody (RP1104).
CACYBP was detected in a paraffin-embedded section of mouse testis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-CACYBP Antibody (RP1104) 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) (Catalog # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of CACYBP using anti-CACYBP antibody (RP1104).
CACYBP was detected in a paraffin-embedded section of rat testis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-CACYBP Antibody (RP1104) 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) (Catalog # SA1022) with DAB as the chromogen.

Figure 4. IHC analysis of CACYBP using anti-CACYBP antibody (RP1104).
CACYBP was detected in a paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-CACYBP Antibody (RP1104) 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) (Catalog # SA1022) with DAB as the chromogen.

Figure 5. ICC analysis of CACYBP using anti-CACYBP antibody (RP1104).
CACYBP was detected in immunocytochemical section of MCF-7 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1μg/ml rabbit anti-CACYBP Antibody (RP1104) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 6. Immunoprecipitating CACYBP in U251 whole cell lysate.
Western blot analysis of CACYBP using anti-CACYBP antibody (RP1104).
Lane 1: U251 whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-CACYBP antibody in U251 whole cell lysate,
Lane 3: anti-CACYBP antibody (2μg) + U251 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-CACYBP antigen affinity purified polyclonal antibody (RP1104) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for CACYBP at approximately 27 kDa. The expected band size for CACYBP is at 27 kDa.