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

製品の概要

  • 製品名Anti-Cytochrome C antibody [7H8.2C12]
    Cytochrome C 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [7H8.2C12] to Cytochrome C
  • アプリケーション適用あり: Flow Cyt, IHC-Fr, ICC/IF, WB, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Horse, Pigeon, Human, Drosophila melanogaster
  • 免疫原

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

  • エピトープThe antibody recognizes an epitope within amino acids 93-104 of pigeon Cytochrome C, based on competitive ELISA results.
  • ポジティブ・コントロール
    • This 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.
  • 特記事項

    Alternative versions available:

    Anti-Cytochrome C antibody (HRP) [7H8.2C12] (ab199222)

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab13575 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Flow Cyt Use 0.1-1µg for 106 cells.

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

IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/200 - 1/500. (see Abreviews)
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 12 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

  • 機能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.
  • 関連疾患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.
  • 配列類似性Belongs to the cytochrome c family.
  • 翻訳後修飾Binds 1 heme group per subunit.
  • 細胞内局在Mitochondrion matrix.
  • Information by UniProt
  • 参照データベース
  • 別名
    • CYC antibody
    • CYC_HUMAN antibody
    • CYCS antibody
    • Cytochrome c antibody
    • Cytochrome c somatic antibody
    • HCS antibody
    • THC4 antibody
    see all

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

  • 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 : Human heart tissue lysate - total protein (ab29431)

    Lysates/proteins at 10 µg per lane.

    Secondary
    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 : 12 kDa
    Additional bands at : 70 kDa. We are unsure as to the identity of these extra bands.

    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.
  • 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.
  • 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

  • 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

  • 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.
  • 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.

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

This product has been referenced in:
  • Zhao X  et al. Effects of 17-AAG on the cell cycle and apoptosis of H446 cells and the associated mechanisms. Mol Med Rep 14:1067-74 (2016). WB ; Human . Read more (PubMed: 27279418) »
  • Barneo-Muñoz M  et al. Lack of GDAP1 induces neuronal calcium and mitochondrial defects in a knockout mouse model of charcot-marie-tooth neuropathy. PLoS Genet 11:e1005115 (2015). Read more (PubMed: 25860513) »

See all 25 Publications for this product

Product Wall

Application Western blot
Sample Rat Tissue lysate - other (Cytosolic Fractions from Skeletal Muscle)
Gel Running Conditions Reduced Denaturing (15%)
Loading amount 20 µg
Specification Cytosolic Fractions from Skeletal Muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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投稿 Feb 18 2016

Application Western blot
Sample Mouse Tissue lysate - other (Cytosolic Fractions from Skeletal Muscle)
Gel Running Conditions Reduced Denaturing (15%)
Loading amount 20 µg
Specification Cytosolic Fractions from Skeletal Muscle
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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投稿 Feb 18 2016

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 25% · Temperature: 25°C
Sample Sheep Cell (brain tissue (cerebellum))
Specification brain tissue (cerebellum)
Permeabilization Yes - 0.1% Triton X-100, 5' RT
Fixative ZSF
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投稿 Sep 01 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (h9c2)
Specification h9c2
Fixative Paraformaldehyde
Permeabilization Yes - 1% triton x100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C
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投稿 Mar 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Liver)
Loading amount 50 µg
Specification Liver
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Verified customer

投稿 Jan 07 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Liver)
Specification Liver
Fixative Acetone
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
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Abcam user community

Verified customer

投稿 Jan 07 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (Liver)
Loading amount 50 µg
Specification Liver
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Dec 03 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Liver)
Specification Liver
Fixative Acetone
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Username

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Verified customer

投稿 Nov 30 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - other (Cytosolic fractions from GTN skeletal muscles)
Loading amount 50 µg
Specification Cytosolic fractions from GTN skeletal muscles
Gel Running Conditions Reduced Denaturing (15% gel)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Dr. G.K Sakellariou

Verified customer

投稿 Sep 28 2012

Thank you for your call today and for letting us know about the trouble with ab13575.
As we discussed, I'm sending a free of charge vial of ab110325 which should arrive on Tuesday.
Please keep me updated about the results with this replacemen...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"