Overview

  • Product nameAnti-Cytochrome C antibody
    See all Cytochrome C primary antibodies
  • Description
    Rabbit polyclonal to Cytochrome C
  • Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Sheep, Rabbit, Horse, Cow, Pig, Non Human Primates
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Cytochrome C.

    (Peptide available as ab100818.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: HeLa Whole Cell; HepG2 Whole Cell; MCF7 Whole Cell; HEK293 Whole Cell; Human Testis Tissue; Human Liver Tissue; Mouse Heart Tissue; Rat Heart Tissue. IHC-P: Human skeletal muscle FFPE tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab90529 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 12 kDa).

Target

  • FunctionElectron carrier protein. The oxidized form of the cytochrome c heme group can accept an electron from the heme group of the cytochrome c1 subunit of cytochrome reductase. Cytochrome c then transfers this electron to the cytochrome oxidase complex, the final protein carrier in the mitochondrial electron-transport chain.
    Plays a role in apoptosis. Suppression of the anti-apoptotic members or activation of the pro-apoptotic members of the Bcl-2 family leads to altered mitochondrial membrane permeability resulting in release of cytochrome c into the cytosol. Binding of cytochrome c to Apaf-1 triggers the activation of caspase-9, which then accelerates apoptosis by activating other caspases.
  • Involvement in diseaseDefects in CYCS are the cause of thrombocytopenia type 4 (THC4) [MIM:612004]; also known as autosomal dominant thrombocytopenia type 4. Thrombocytopenia is the presence of relatively few platelets in blood. THC4 is a non-syndromic form of thrombocytopenia. Clinical manifestations of thrombocytopenia are absent or mild. THC4 may be caused by dysregulated platelet formation.
  • Sequence similaritiesBelongs to the cytochrome c family.
  • Post-translational
    modifications
    Binds 1 heme group per subunit.
  • Cellular localizationMitochondrion matrix.
  • Information by UniProt
  • Database links
  • Alternative names
    • CYC antibody
    • CYC_HUMAN antibody
    • CYCS antibody
    • Cytochrome c antibody
    • Cytochrome c somatic antibody
    • HCS antibody
    • THC4 antibody
    see all

Anti-Cytochrome C antibody images

  • ab90529 staining cytochrome C in HepG2 cells treated with mevastatin sodium salt (ab120854), by ICC/IF. Increase of cytochrome C expression correlates with increased concentration of mevastatin sodium salt, as described in literature.
    The cells were incubated at 37°C for 4 hours in media containing different concentrations of ab120854 (mevastatin sodium salt) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab90529 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • ab90529 staining cytochrome C in HepG2 cells treated with atorvastatin hemicalcium salt (ab141083), by ICC/IF. Increase of cytochrome C expression correlates with increased concentration of atorvastatin hemicalcium salt, as described in literature.
    The cells were incubated at 37°C for 4 hours in media containing different concentrations of ab141083 (atorvastatin hemicalcium salt) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab90529 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • ab90529 staining cytochrome C in HepG2 cells treated with mevastatin (ab120652), by ICC/IF. Increase of cytochrome C expression correlates with increased concentration of mevastatin, as described in literature.
    The cells were incubated at 37°C for 4 hours in media containing different concentrations of ab120652 (mevastatin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab90529 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • ab90529 staining cytochrome C in HepG2 cells treated with fluvastatin sodium salt (ab120651), by ICC/IF. Increase of cytochrome C expression correlates with increased concentration of fluvastatin sodium salt, as described in literature.
    The cells were incubated at 37°C for 48 hours in media containing different concentrations of ab120651 (fluvastatin sodium salt) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab90529 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • All lanes : Anti-Cytochrome C antibody (ab90529) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : Human testis tissue lysate - total protein (ab30257)
    Lane 6 : Human liver tissue lysate - total protein (ab29889)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 12 kDa
    Observed band size : 15 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 minutes
  • ICC/IF image of ab90529 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab90529 at 1µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in HeLa, Hek293, and MCF-7 formaldehyde (4%) (10min) fixed cells.

  • All lanes : Anti-Cytochrome C antibody (ab90529) at 1 µg/ml

    Lane 1 : Heart (Mouse) Tissue Lysate
    Lane 2 : Heart (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 12 kDa
    Observed band size : 14 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 30 kDa,65 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes
  • IHC image of Cytochrome C staining in human skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab90529, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-Cytochrome C antibody (ab90529)

This product has been referenced in:
  • Zhong R  et al. Kuntai Capsule Inhibited Endometriosis via Inducing Apoptosis in a Rat Model. Evid Based Complement Alternat Med 2016:5649169 (2016). Rat . Read more (PubMed: 27597876) »
  • Kannan A  et al. Mitochondrial Reprogramming Regulates Breast Cancer Progression. Clin Cancer Res 22:3348-60 (2016). Read more (PubMed: 26888829) »

See all 5 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (Epithelial OVCA cell line)
Gel Running Conditions Reduced Denaturing (12.5)
Loading amount 100 µg
Treatment 0- 200µM Perifosine 24 hr
Specification Epithelial OVCA cell line
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Aug 09 2016

Application Western blot
Sample Rat Tissue lysate - whole (Rat Gonad)
Gel Running Conditions Reduced Denaturing (12.5%)
Loading amount 75 µg
Specification Rat Gonad
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Nov 18 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Gonad)
Antigen retrieval step Enzymatic
Permeabilization No
Specification Gonad
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Sep 30 2015

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