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).

Reviews (2) Submit a question References (72)

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.
    Database link: P99999

  • Positive control

    • WB: Human fetal kidney and fetal heart tissue lysate. Rat and mouse brain tissue lysate. Molt4 and SH-SY5Y cell lysate. Human heart, kidney and spleen lysate. ICC/IF: SH-SY5Y and HeLa cells. IHC-P: Human cervical carcinoma tissue. Mouse liver tissue. Human and rat kidney tissue. IP: Molt-4 cells.

  • General notes

    A trial size is available to purchase for this antibody.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

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

    see all

  • Alternative names

    • CYC antibody
    • CYC_HUMAN antibody
    • CYCS antibody
    • Cytochrome c antibody
    • Cytochrome c somatic antibody
    • HCS antibody
    • THC4 antibody
    see all

Images

  • Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome C antibody [EPR1327] (ab133504)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome C antibody [EPR1327] (ab133504)

    Immunofluorescent analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labelling Cytochrome C with unpurified ab133504 at 1/100 dilution.

  • Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    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

  • Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    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

  • Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (ab133504)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (ab133504)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (ab133504)

    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.

  • Immunoprecipitation - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Immunoprecipitation - Anti-Cytochrome C antibody [EPR1327] (ab133504)

    ab133504 (purified) at 1/30 immunoprecipitating Cytochrome C in Molt-4 cells.

    For western blotting, a HRP-conjugated goat anti-rabbit (H+L), was used as the secondary antibody (1/1000).

    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

  • Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Western blot - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    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?

  • Other - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Other - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (ab133504)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody [EPR1327] (ab133504)

    Immunohistochemical analysis of paraffin-embedded human kidney tissue labelling Cytochrome C with unpurified ab133504 at 1/250 dilution.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Liu L  et al. CP-31398 attenuates endometrial cancer cell invasion, metastasis and resistance to apoptosis by downregulating MDM2 expression. Int J Oncol 54:942-954 (2019). Read more (PubMed: 30628640) »
  • Wang L  et al. The Effect of Overexpression of the Enhancer of Zeste Homolog 1 (EZH1) Gene on Aristolochic Acid-Induced Injury in HK-2 Human Kidney Proximal Tubule Cells In Vitro. Med Sci Monit 25:801-810 (2019). Read more (PubMed: 30688289) »
See all 72 Publications for this product

Publishing research using ab133504? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-Cytochrome C antibody [EPR1327]

 Good
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted May 17 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-Cytochrome C antibody [EPR1327]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

Submitted Oct 06 2017

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