Product Name
Cytochrome C Recombinant Rabbit Monoclonal Antibody [SC69-08] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human Cytochrome C aa 20-60.
Validated Applications
WB, IF-Tissue, IHC-P, IP, IF-Cell
Target Molecular Weight
Predicted band size: 12 kDa
Positive Control
HeLa cell lysate, C6 cell lysate, Mouse brain tissue lysate, Mouse kidney tissue lysate, Rat brain tissue lysate, Rat kidney tissue lysate, Jurkat cell lysate, Human kidney tissue lysate, C6, human kidney tissue, mouse liver tissue, mouse kidney tissue, rat brain tissue.
Storage Instructions
Store at 2-8℃. Avoid freeze.
Purification Method
Protein A affinity purified.
Function
Cytochrome c is a well characterized mobile electron transport protein that is essential to energy conversion in all aerobic organisms. In mammalian cells, this highly conserved protein is normally localized to the mitochondrial intermembrane space. More recent studies have identifed cytosolic cytochrome c as a factor necessary for activation of apoptosis. During apoptosis, cytochrome c is translocated from the mitochondrial membrane to the cytosol, where it is required for activation of caspase-3 (CPP32). Overexpression of Bcl-2 has been shown to prevent the translocation of cytochrome c, thereby blocking the apoptotic process. Overexpression of Bax has been shown to induce the release of cytochrome c and to induce cell death. The release of cytochrome c from the mitochondria is thought to trigger an apoptotic cascade, whereby Apaf-1 binds to Apaf-3 (caspase-9) in a cytochrome c-dependent manner, leading to caspase-9 cleavage of caspase-3.
Background References
1. Liu Z et al. Mark4 promotes oxidative stress and inflammation via binding to PPAR and activating NF-kB pathway in mice adipocytes. Sci Rep 6:21382 (2016).
2. Qi Y et al. PGC-1a Silencing Compounds the Perturbation of Mitochondrial Function Caused by Mutant SOD1 in Skeletal Muscle of ALS Mouse Model. Front Aging Neurosci 7:204 (2015).
Sequence Similarity
Belongs to the cytochrome c family.
Post-translational Modification
Binds 1 heme group per subunit.; Phosphorylation at Tyr-49 and Tyr-98 both reduce by half the turnover in the reaction with cytochrome c oxidase, down-regulating mitochondrial respiration.
Subcellular Location
Mitochondrion intermembrane space.
Synonyms
CYC antibody
CYC_HUMAN antibody
CYCS antibody
Cytochrome c antibody
Cytochrome c somatic antibody
HCS antibody
THC4 antibody
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This data was developed using ET1610-60, the same antibody clone in a different buffer formulation.
Western blot analysis of Cytochrome C on different lysates with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/2,000 dilution.
Lane 1: HeLa cell lysate (10 µg/Lane)
Lane 2: C6 cell lysate (10 µg/Lane)
Lane 3: Mouse brain tissue lysate (20 µg/Lane)
Lane 4: Mouse kidney tissue lysate (20 µg/Lane)
Lane 5: Rat brain tissue lysate (20 µg/Lane)
Lane 6: Rat kidney tissue lysate (20 µg/Lane)
Predicted band size: 12 kDa
Observed band size: 12 kDa
Exposure time: 4 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-60) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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This data was developed using ET1610-60, the same antibody clone in a different buffer formulation.
Western blot analysis of Cytochrome C on different lysates with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/2,000 dilution.
Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: Jurkat cell lysate (15 µg/Lane)
Lane 3: C6 cell lysate (15 µg/Lane)
Lane 4: Human kidney tissue lysate (20 µg/Lane)
Lane 5: Mouse brain tissue lysate (20 µg/Lane)
Lane 6: Rat brain tissue lysate (20 µg/Lane)
Predicted band size: 12 kDa
Observed band size: 12 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-60) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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This data was developed using ET1610-60, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of C6 cells labeling Cytochrome C with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
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This data was developed using ET1610-60, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-60) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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This data was developed using ET1610-60, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-60) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
This data was developed using ET1610-60, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-60) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
This data was developed using ET1610-60, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Cytochrome C antibody (ET1610-60) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-60) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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