Recombinant
RabMAb

Recombinant Anti-MTCO2 antibody [EPR3314] (ab79393)

Overview

  • Product name

    Anti-MTCO2 antibody [EPR3314]
    See all MTCO2 primary antibodies
  • Description

    Rabbit monoclonal [EPR3314] to MTCO2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human MTCO2 aa 200-300. The exact sequence is proprietary.

  • Positive control

    • K562, MCF7, THP1 and HeLa cell lysates; human heart, liver and heart muscle tissues.
  • General notes

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab79393 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
ICC/IF Use a concentration of 5 µg/ml.
WB 1/5000 - 1/10000. Detects a band of approximately 21 kDa (predicted molecular weight: 25 kDa).
IP 1/30.
IHC-P 1/100 - 1/250. Antigen retrieval is not essential but may optimise staining.
Flow Cyt 1/60.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Cytochrome c oxidase is the component of the respiratory chain that catalyzes the reduction of oxygen to water. Subunits 1-3 form the functional core of the enzyme complex. Subunit 2 transfers the electrons from cytochrome c via its binuclear copper A center to the bimetallic center of the catalytic subunit 1.
  • Involvement in disease

    Defects in MT-CO2 are a cause of mitochondrial complex IV deficiency (MT-C4D) [MIM:220110]; also known as cytochrome c oxidase deficiency. A disorder of the mitochondrial respiratory chain with heterogeneous clinical manifestations, ranging from isolated myopathy to severe multisystem disease affecting several tissues and organs. Features include hypertrophic cardiomyopathy, hepatomegaly and liver dysfunction, hypotonia, muscle weakness, excercise intolerance, developmental delay, delayed motor development and mental retardation. A subset of patients manifest Leigh syndrome.
  • Sequence similarities

    Belongs to the cytochrome c oxidase subunit 2 family.
  • Cellular localization

    Mitochondrion inner membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • COII antibody
    • COX 2 antibody
    • COX II antibody
    • COX2 antibody
    • COX2_HUMAN antibody
    • COXII antibody
    • Cytochrome c oxidase II antibody
    • Cytochrome c oxidase polypeptide II antibody
    • Cytochrome c oxidase subunit 2 antibody
    • MT CO2 antibody
    • MT-CO2 antibody
    • MTCO2 antibody
    see all

Images

  • All lanes : Anti-MTCO2 antibody [EPR3314] (ab79393) at 1/10000 dilution

    Lane 1 : K562 cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : THP1 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 25 kDa
    Observed band size: 21 kDa
    why is the actual band size different from the predicted?

  • ab79393 at 1/100 dilution staining Cytochrome C oxidase subunit II in Human Liver (A), Human Heart(B) and HeLa Cell Line(C) by Immunohistochemistry, Paraffin-embedded tissues. Detection method of A and B was HRP-conjugated anti-rabbit with DAB substrate used for staining.

  • ICC/IF image of ab79393 stained HepG2 cells. The cells were 100% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79393, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HepG2 cells stained with ab79393 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab79393, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:

  • Xian H  et al. STX17 dynamically regulated by Fis1 induces mitophagy via hierarchical macroautophagic mechanism. Nat Commun 10:2059 (2019). Read more (PubMed: 31053718) »
  • Balboa D  et al. Insulin mutations impair beta-cell development in a patient-derived iPSC model of neonatal diabetes. Elife 7:N/A (2018). Read more (PubMed: 30412052) »
See all 15 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Question

I would like to know if the epitopes recognized by the following antibodies are known:
Code
Name
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-12C4F12--ab110258.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-12C4F12--ab110258.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-12C4F12--ab110258.html
Mouse monoclonal
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-EPR3314-ab79393.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-EPR3314-ab79393.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-EPR3314-ab79393.html
Rabbit monoclonal
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-4B12A5-ab110271.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-4B12A5-ab110271.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-4B12A5-ab110271.html
Mouse monoclonal
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-ab91317.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-ab91317.html
Rabbit polyclonal
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-EPR3313-ab109739.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-EPR3313-ab109739.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-EPR3313-ab109739.html
Rabbit monoclonal
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-ab101802.html
https://www.abcam.com/Cytochrome-C-oxidase-subunit-II-antibody-ab101802.html
Rabbit polyclonal
Awaiting your kind reply, I remain

Read More
Answer

Thank you for contacting us.

The immunogens are as follows;

ab79393; A synthetic peptide corresponding to residues in human Cytochrome C oxidase subunit II
ab109739; A synthetic peptide corresponding to residues in Human Cytochrome C oxidase subunit II.
ab101802; A synthetic peptide corresponding to residues in Human Cytochrome C oxidase subunit II.
ab91317; A synthetic peptide corresponding to residues in Human Cytochrome C oxidase subunit II.
ab110258: exact epitope is not determined. Antigen was full length protein
ab110271: exact epitope is not determined. Antigen was full length protein

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa WT vs HeLa rho-0)
Loading amount
13 µg
Specification
HeLa WT vs HeLa rho-0
Gel Running Conditions
Reduced Denaturing (Reduced Denaturing (4-15%))
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jul 31 2012

Answer

Thank you for contacting Abcam.


For IHC and ICC, antibodies are typically diluted in PBS or in a 1-3% BSA in PBS solution. Some researchers will use TBS instead, but either is fine. I have included links below to our IHC and ICC protocols for your reference.


https://www.abcam.com/index.html?pageconfig=resource&rid=11384


https://www.abcam.com/index.html?pageconfig=resource&rid=11417


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

Read More

Answer

Thank you for your enquiry.

I have confirmed that ab79393, anti-Cytochrome C oxidase subunit II antibody antibody is sold as tissue culture supernatant. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for tissue culture supernatant, concentration of antibody is known to very between 1 - 3 mg/ml.

While we do not offer a product with catalog number ab25938 our Rabbit IgG, monoclonal [SP137] - isotype control with designation ab128142. We recommend using this product at a dilution of 1:100 for IHC-P control.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Question
Answer

The immunogen of ab79393 shares 70% identity with rat Cytochrome c oxidase subunit 2. It also shares 70% identity with cat. Based on these number I would not predict cross-reactivity with either species.I have looked through the other antibodies in our catalogue and I do not think that they will react either cat or rat tissue.

Read More
Application
Western blot
Sample
Human Cell lysate - whole cell (PBMC)
Loading amount
20 µg
Specification
PBMC
Gel Running Conditions
Reduced Denaturing
Blocking step
Fluorescence special buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 04 2011

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