Name: Anti-Cytochrome C antibody [EPR1327]
Brand: RabMAb
SKU: ab133504
Rating: 5 (2 reviews)

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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Dissociation constant (K_D)

K_D = 1.29 x 10 ^-10 M
Learn more about K_D
Storage buffer

pH: 7.40
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR1327
Isotype

IgG
Research areas

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.

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.
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: P99999 Human
- SwissProt: P62897 Mouse
- SwissProt: P62898 Rat
- Unigene: 437060 Human
- Unigene: 617193 Human
- Unigene: 35389 Mouse
- Unigene: 2202 Rat
see all
Alternative names
- CYC antibody
- CYC_HUMAN antibody
- CYCS antibody
- Cytochrome c antibody
- Cytochrome c somatic antibody
- HCS antibody
- THC4 antibody
see all

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)

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)

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

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)

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)

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)

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)

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)

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.
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)

Equilibrium disassociation constant (K_D)
Learn more about K_D

Click here to learn more about K_D
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.

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

Yan H et al. NR4A1-induced increase in the sensitivity of a human gastric cancer line to TNFa-mediated apoptosis is associated with the inhibition of JNK/Parkin-dependent mitophagy. Int J Oncol 52:367-378 (2018). Read more (PubMed: 29207128) »
Sun D et al. bFGF plays a neuroprotective role by suppressing excessive autophagy and apoptosis after transient global cerebral ischemia in rats. Cell Death Dis 9:172 (2018). WB ; Rat . Read more (PubMed: 29416039) »

See all 39 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

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

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

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Anti-Cytochrome C antibody [EPR1327] (ab133504)

Overview

Properties

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Images

Protocols

Datasheets and documents

References

This product has been referenced in:

Customer reviews and Q&As

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

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