Overview

  • Product name

    Anti-Cytochrome C antibody [7H8.2C12] - BSA and Azide free
    See all Cytochrome C primary antibodies
  • Description

    Mouse monoclonal [7H8.2C12] to Cytochrome C - BSA and Azide free
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, IHC-P, IHC-Fr, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    corresponding to Cytochrome C.

  • Epitope

    The antibody recognizes an epitope within amino acids 93-104 of pigeon Cytochrome C, based on competitive ELISA results.
  • Positive control

    • WB: HeLa, Jurkat and human heart whole cell lysates; IHC-P: Human liver and skin tissues; ICC/IF: Leukocytes from murine bone marrow; FC: HepG2 cells.
  • General notes

    Ab237966 is a PBS only version of ab13575.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Applications

Our Abpromise guarantee covers the use of ab237966 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
WB Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 11 kDa.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 0.1-1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ICC/IF 1/200 - 1/500.

Target

  • Function

    Electron 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 disease

    Defects 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 similarities

    Belongs to the cytochrome c family.
  • Post-translational
    modifications

    Binds 1 heme group per subunit.
  • Cellular localization

    Mitochondrion 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

Images

  • All lanes : Anti-Cytochrome C antibody [7H8.2C12] (ab13575) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : ab29431

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 11 kDa
    Observed band size: 14 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Abcam recommends using milk (5%) as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab13575).

  • IHC image of Cytochrome C staining in human normal liver 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 ab13575, 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab13575).

  • ab13575 staining Cytochrome C in leukocytes from murine bone marrow by Immunocytochemistry/ Immunofluorescence. The cells were fixed in methanol and then blocked using 5% serum for 2 hours at 25°C. Samples were then incubated with primary antibody at 1/250 for 16 hours at 5°C. The secondary antibody used was a goat anti-mouse IgG conjugated to Alexa Fluor® 594 (red) used at a 1/500 dilution. Counterstained with DAPI (blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab13575).

  • Overlay histogram showing HepG2 cells stained with ab13575 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13575, 0.1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1]/mouse IgG2b [PLPV219] (ab91353/ab91366, 1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab13575).

  • Ab13575 staining human normal skin tissue. Staining is localised to mitochondria.
    Left panel: with primary antibody at 4 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab13575).

References

ab237966 has not yet been referenced specifically in any publications.

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