Anti-Cytochrome C [7H8.2C12] antibody (ab13575)

Overview

• Product nameAnti-Cytochrome C [7H8.2C12] antibodySee all Cytochrome C primary antibodies ...
• Description
  Mouse monoclonal [7H8.2C12] to Cytochrome C
• Tested applicationsFlow Cyt, IHC-Fr, ICC/IF, WB, IHC-P more details
• Species reactivity
  Reacts with: Mouse, Rat, Horse, Pigeon, Human, Fruit fly (Drosophila melanogaster)
  
• Immunogen

  Synthetic peptides corresponding to amino acids 1-80, 81-104 and 66-104 of pigeon CYT.

• EpitopeThe antibody recognizes an epitope within amino acids 93-104 of pigeon Cytochrome C, based on competitive ELISA results.
• Positive controlThis antibody gave a positive signal in the following lysates: HeLa whole cell; Jurkat whole cell; Human heart tissue. In Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised HepG2 cells.

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: PBS, pH 7.4
• Concentration information loading...
• PurityIgG fraction
• ClonalityMonoclonal
• Clone number7H8.2C12
• IsotypeIgG
• Research Areas
  • Metabolism
  • Types of disease
  • Cancer

  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Oxidative phosphorylation

  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Mitochondrial markers

  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Cytochromes

  • Metabolism
  • Pathways and Processes
  • Metabolic signaling pathways
  • Lipid and lipoprotein metabolism
  • Lipases

  • Metabolism
  • Pathways and Processes
  • Metabolism processes
  • Apoptosis

  • Metabolism
  • Pathways and Processes
  • Metabolic signaling pathways
  • Energy transfer pathways
  • Integration of energy

  • Metabolism
  • Pathways and Processes
  • Metabolic signaling pathways
  • Energy transfer pathways
  • Energy Metabolism

  • Cancer
  • Invasion/microenvironment
  • Apoptosis
  • Cytochrome C

  • Cancer
  • Cancer Metabolism
  • Metabolic signaling pathway
  • Integration of energy metabolism

  • Signal Transduction
  • Metabolism
  • Mitochondrial

  • Signal Transduction
  • Metabolism
  • Energy Metabolism

  • Cardiovascular
  • Lipids / Lipoproteins
  • Lipid Metabolism
  • Cytochromes

  • Cell Biology
  • Apoptosis
  • Mitochondrial

Applications

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
  modificationsBinds 1 heme group per subunit.
• Cellular localizationMitochondrion matrix.

Target information above from: UniProt accession P99999 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 54205 Human
  • Entrez Gene: 13063 Mouse
  • Entrez Gene: 25309 Rat
  • Omim: 123970 Human
  • SwissProt: P00004 Horse
  • SwissProt: P99999 Human
  • SwissProt: P62897 Mouse
  • SwissProt: P62898 Rat

  • Unigene: 437060 Human
  • Unigene: 617193 Human
  • Unigene: 2202 Rat

  see all

• Alternative names
  CYC antibodyCYC_HUMAN antibodyCYCS antibody

  Cytochrome c antibodyCytochrome c somatic antibodyHCS antibodyTHC4 antibody

  see all

Anti-Cytochrome C [7H8.2C12] antibody images

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Cytochrome C antibody [7H8.2C12] (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.
• 

  Immunohistochemistry (Frozen sections) - Cytochrome C antibody [7H8.2C12] (ab13575)This image is courtesy of an anonymous Abreview
  ab13575 staining Cytochrome C in mouse liver tissue sections by IHC-Fr (formaldehyde-fixed frozen sections). Tissue samples were fixed with formaldehyde and blocked with 2% BSA for 30 minutes at 20°C. The sample was incubated with primary antibody (1/200 in PBS) at 4°C for 9 hours. An Alexa Fluor^®555-conjugated Goat polyclonal to mouse IgG (1/200) was used as secondary antibody. Nuclei were stained with DAPI.

  See Abreview

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Cytochrome C antibody [7H8.2C12] (ab13575)
  IHC image of Cytochrome C staining in human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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.
• 

  Immunocytochemistry/ Immunofluorescence - Cytochrome C antibody [7H8.2C12] (ab13575)Image courtesy of an anonymous Abreview.
  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).

  See Abreview

• 

  Western blot - Anti-Cytochrome C antibody [7H8.2C12] (ab13575)
  All lanes : Anti-Cytochrome C [7H8.2C12] antibody (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 : Heart (Human) Tissue Lysate - adult normal tissue (ab29431)
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (ab97040) at 1/5000 dilution
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 12 kDa
  Additional bands at : 70 kDa. We are unsure as to the identity of these extra bands.
  
  Exposure time : 3 minutes
• 

  Flow Cytometry-Anti-Cytochrome C antibody [7H8.2C12](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, 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 IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.