Anti-Cytochrome C/Cycs Antibody

• Catalog Number: orb402533
• Category: Antibodies
• Description: Anti-Cytochrome C/Cycs Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: IHC, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E. coli-derived mouse Cytochrome C recombinant protein (Position: G2-E105). Mouse Cytochrome C shares 91.3% and 100% amino acid (aa) sequence identity with human and rat Cytochrome C, respectively.
• 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: 14 kDa
• UniProt ID: P62897
• 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: Cytochrome c, somatic; Cycs
• Note: For research use only
• Application notes: Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

IHC analysis of Cytochrome C using anti-Cytochrome C antibody. Cytochrome C was detected in paraffin-embedded section of mouse brain tissue. 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 1 µg/ml rabbit anti-Cytochrome C 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 Cytochrome C using anti-Cytochrome C antibody. Cytochrome C was detected in paraffin-embedded section of mouse intestine tissue. 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 1 µg/ml rabbit anti-Cytochrome C 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 Cytochrome C using anti-Cytochrome C antibody. Cytochrome C was detected in paraffin-embedded section of rat brain tissue. 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 1 µg/ml rabbit anti-Cytochrome C 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 Cytochrome C using anti-Cytochrome C antibody. Cytochrome C was detected in paraffin-embedded section of rat intestine tissue. 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 1 µg/ml rabbit anti-Cytochrome C 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 Cytochrome C using anti-Cytochrome C 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: human HL-60 whole cell lysates, Lane 2: human K562 whole cell lysates, Lane 3: human U-937 whole cell lysates, Lane 4: human U20S whole cell lysates, Lane 5: human T-47D whole cell lysates, Lane 6: human PC-3 whole cell lysates, Lane 7: human CACO-2 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-Cytochrome C 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 Cytochrome C at approximately 14 kDa. The expected band size for Cytochrome C is at 12 kDa.

Western blot analysis of Cytochrome C using anti-Cytochrome C 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 brain tissue lysates, Lane 2: rat heart tissue lysates, Lane 3: rat kidney tissue lysates, Lane 4: rat C6 whole cell lysates, Lane 5: mouse brain tissue lysates, Lane 6: mouse heart tissue lysates, Lane 7: mouse kidney tissue lysates, Lane 8: mouse Neuro-2a 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-Cytochrome C 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 Cytochrome C at approximately 14 kDa. The expected band size for Cytochrome C is at 12 kDa.