Anti-Cytochrome C/CYCS Antibody Picoband®

Figure 1. Western blot analysis of Cytochrome c/CYCS using anti-Cytochrome c/CYCS antibody (PA1118).
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 Hela whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human HT1080 whole cell lysates,
Lane 5: rat kidney tissue lysates,
Lane 6: rat skeletal muscle tissue lysates,
Lane 7: mouse kidney tissue lysates,
Lane 8: mouse skeletal muscle 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-Cytochrome c/CYCS antigen affinity purified polyclonal antibody (PA1118) 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 Cytochrome c/CYCS at approximately 14 kDa. The expected band size for Cytochrome c/CYCS is at 12 kDa.

Anti-Cytochrome C antibody, PA1118, IHC(P)
IHC(P): Rat Lung Tissue

Anti-Cytochrome C antibody, PA1118, IHC(F)
IHC(F): Rat Brain Tissue

Anti-Cytochrome C antibody, PA1118, ICC
ICC: A549 Cell

Figure 5. IHC analysis of Cytochrome C using anti-Cytochrome C antibody (PA1118).
Cytochrome C was detected in paraffin-embedded section of human skeletal muscle tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Cytochrome C Antibody (PA1118) 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 6. IHC analysis of Cytochrome C using anti-Cytochrome C antibody (PA1118).
Cytochrome C was detected in paraffin-embedded section of rat kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Cytochrome C Antibody (PA1118) 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 7. IF analysis of CYCS using anti-CYCS antibody (PA1118).
CYCS was detected in immunocytochemical section of A431 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 2μg/mL rabbit anti-CYCS Antibody (PA1118) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.

Figure 8. Flow Cytometry analysis of CACO-2 cells using anti-CYCS antibody (PA1118).
Overlay histogram showing CACO-2 cells stained with PA1118 (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-CYCS Antibody (PA1118, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.