Recombinant
RabMAb

Recombinant Anti-Cytochrome C antibody [EPR1327] (ab133504)

Rabbit recombinant monoclonal Cytochrome C antibody [EPR1327]. Validated in WB, IP, IHC, ICC/IF and tested in Mouse, Rat, Human. Cited in 39 publication(s). Independently reviewed in 2 review(s).

Overview

  • Product name
    Anti-Cytochrome C antibody [EPR1327]
    See all Cytochrome C primary antibodies
  • Description
    Rabbit monoclonal [EPR1327] to Cytochrome C
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Cytochrome C aa 1-100. The exact sequence is proprietary.

  • Positive control
    • Molt4, SH-SY5Y, Human heart, Human kidney and Human spleen lysates. Human kidney tissue.
  • General notes

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133504 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 1/5000. Detects a band of approximately 14 kDa (predicted molecular weight: 11 kDa).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

ICC/IF 1/100.
IP 1/30.

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

  • Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/5000 dilution (purified) + Human fetal kidney tissue lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunofluorescence staining of SH-SY5Y cells with purified ab133504 at a working dilution of 1 in 100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti rabbit (ab150077), used at a dilution of 1 in 500. ab7291 was used to stain tubulin, and this is shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom middle and right hand panels - for the negative controls, purified ab133504 was used at a dilution of 1/200 followed by an Alexa Fluor® 594 goat anti-mouse antibody at a dilution of 1/500.

  • Immunofluorescent analysis of HeLa cells labelling Cytochrome C with unpurified ab133504 at 1/100 dilution.

  • Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/5000 dilution (purified) + Human fetal heart tissue lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/5000 dilution (purified) + Rat brain tissue lysate at 1/1000 dilution

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/50000 dilution (purified) + Mouse brain tissue lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab133504 at a working dilution of 1 in 500. The secondary antibody used is a HRP goat anti-rabbit H+L (ab97051). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded mouse liver with purified ab133504 at a working dilution of 1 in 500. The secondary antibody used is a HRP goat anti-rabbit H+L (ab97051). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded rat kidney with purified ab133504 at a working dilution of 1 in 500. The secondary antibody used is a HRP goat anti-rabbit H+L (ab97051). The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • ab133504 (purified) at 1/30 immunoprecipitating Cytochrome C in Molt-4 cells (Lane 1). For western blotting, a HRP-conjugated goat anti-rabbit (H+L), was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-Cytochrome C antibody [EPR1327] (ab133504) at 1/10000 dilution (unpurified)

    Lane 1 : Molt4 lysate
    Lane 2 : SH-SY5Y lysate
    Lane 3 : Human heart lysate
    Lane 4 : Human kidney lysate
    Lane 5 : Human spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat-anti-rabbit HRP at 1/2000 dilution

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

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling Cytochrome C with unpurified ab133504 at 1/250 dilution.

References

This product has been referenced in:
  • Le L  et al. The protective effects of the native flavanone flavanomarein on neuronal cells damaged by 6-OHDA. Phytomedicine 53:193-204 (2019). Read more (PubMed: 30668399) »
  • Wang Q  et al. Efficacy of celastrol combined with cisplatin in enhancing the apoptosis of U-2OS osteosarcoma cells via the mitochondrial and endoplasmic reticulum pathways of apoptosis. Oncol Lett 17:3305-3313 (2019). Read more (PubMed: 30867764) »
See all 41 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Rabbit Tissue lysate - other (cardiomyocytes)
Gel Running Conditions
Reduced Denaturing (gel 12%)
Loading amount
60 µg
Specification
cardiomyocytes
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted May 17 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human non-small cell lung carcinoma A549)
Permeabilization
Yes - 0.2% Triton X-100
Specification
human non-small cell lung carcinoma A549
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 06 2017

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