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

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR1327] to Cytochrome C
Suitable for: WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Human

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 & Species

Application	Species
ICC/IF	Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

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
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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Immunogen affinity purified
Clonality

Monoclonal
Clone number

EPR1327
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- MOLT-4 whole cell lysate (ab7912)
Recombinant Protein
- Recombinant Human Cytochrome C protein (ab172817)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
ICC/IF	Human
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat Human

Application	Abreviews	Notes
WB	(1)	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)	1/100.
IP		1/30.

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 (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)

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.

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)

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?
OI-RD Scanning - 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.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-Cytochrome C antibody [EPR1327] (ab133504)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

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

ab133504 has been referenced in 102 publications.

Li J et al. Effect of SOD2 methylation on mitochondrial DNA4834-bp deletion mutation in marginal cells under oxidative stress. Bosn J Basic Med Sci 20:70-77 (2020). PubMed: 31465718
Wu H et al. LncRNA ACART protects cardiomyocytes from apoptosis by activating PPAR-?/Bcl-2 pathway. J Cell Mol Med 24:737-746 (2020). PubMed: 31749326
Duan C et al. Drp1 regulates mitochondrial dysfunction and dysregulated metabolism in ischemic injury via Clec16a-, BAX-, and GSH- pathways. Cell Death Dis 11:251 (2020). PubMed: 32312970
Kang L et al. The mitochondria-targeted anti-oxidant MitoQ protects against intervertebral disc degeneration by ameliorating mitochondrial dysfunction and redox imbalance. Cell Prolif 53:e12779 (2020). PubMed: 32020711
Liu H et al. Copper Induces Oxidative Stress and Apoptosis in the Mouse Liver. Oxid Med Cell Longev 2020:1359164 (2020). PubMed: 32411316

View all Publications for this product

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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. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Dissociation constant (KD)

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (102)

Customer reviews and Q&As

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

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

Dissociation constant (K_D)

Post-translational
modifications