MitoSciences (MS404)

Anti-MTCO1 antibody [1D6E1A8] (ab14705)

Overview

  • Product nameAnti-MTCO1 antibody [1D6E1A8]See all MTCO1 primary antibodies ...
  • Description
    Mouse monoclonal [1D6E1A8] to MTCO1
  • SpecificityIn mouse liver lysate a specific band below 37 kDa was detected.
  • Tested applicationsICC/IF, IHC-FoFr, IHC-P, WB, ICC, Flow Cyt, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, Caenorhabditis elegans, Zebrafish, Rhesus monkey, Chinese Hamster
  • Immunogen

    Purified mitochondrial Complex IV subunit I (Human).

  • Positive control
    • Human heart mitochondria. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal colon. In Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised HEK293 cells.
  • General notes

    Samples that are to be used in Western blotting with this antibody should NOT be boiled before loading onto the gel. Boiling of the sample will cause a loss of signal in Western blotting (see WB image on ab110413).

Properties

Applications

Our Abpromise guarantee covers the use of ab14705 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 at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration. PubMed: 23840470
IHC-P Use a concentration of 5 µg/ml.
WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 57 kDa).
ICC Use a concentration of 5 µg/ml.
Flow Cyt Use 1µg for 106 cells.
IHC-Fr Use at an assay dependent concentration.

Target

  • FunctionCytochrome 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. CO I is the catalytic subunit of the enzyme. Electrons originating in cytochrome c are transferred via the copper A center of subunit 2 and heme A of subunit 1 to the bimetallic center formed by heme A3 and copper B.
  • PathwayEnergy metabolism; oxidative phosphorylation.
  • Involvement in diseaseDefects in MT-CO1 are a cause of Leber hereditary optic neuropathy (LHON) [MIM:535000]. LHON is a maternally inherited disease resulting in acute or subacute loss of central vision, due to optic nerve dysfunction. Cardiac conduction defects and neurological defects have also been described in some patients. LHON results from primary mitochondrial DNA mutations affecting the respiratory chain complexes.
    Defects in MT-CO1 are a cause of anemia sideroblastic acquired idiopathic (AISA) [MIM:516030]; a disease characterized by inadequate formation of heme and excessive accumulation of iron in mitochondria.
    Defects in MT-CO1 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.
    Defects in MT-CO1 are associated with recurrent myoglobinuria mitochondrial (RM-MT) [MIM:550500]. Recurrent myoglobinuria is characterized by recurrent attacks of rhabdomyolysis (necrosis or disintegration of skeletal muscle) associated with muscle pain and weakness, and followed by excretion of myoglobin in the urine.
    Defects in MT-CO1 are a cause of deafness sensorineural mitochondrial (DFNM) [MIM:500008]. DFNM is a form of non-syndromic deafness with maternal inheritance. Affected individuals manifest progressive, postlingual, sensorineural hearing loss involving high frequencies.
    Defects in MT-CO1 are a cause of colorectal cancer (CRC) [MIM:114500].
  • Sequence similaritiesBelongs to the heme-copper respiratory oxidase family.
  • Cellular localizationMitochondrion inner membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • COI antibody
    • COX I antibody
    • COX1_HUMAN antibody
    • COXI antibody
    • Cytochrome c oxidase polypeptide I antibody
    • Cytochrome c oxidase subunit 1 antibody
    • Cytochrome C Oxidase subunit I antibody
    • Mitochondrially encoded cytochrome c oxidase I antibody
    • MT CO1 antibody
    • MT-CO1 antibody
    • MTCO 1 antibody
    • MTCO1 antibody
    see all

Anti-MTCO1 antibody [1D6E1A8] images

  • ab14705 staining MTCO1 - Mitochondrial Marker in pig retinal pigment epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% TX-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/2000 in 1% goat serum, 0.1% TX100; PBS) for 16 hours at 4°C. An Alexa Fluor®546-conjugated Goat polyclonal to mouse IgG, dilution 1/500, was used as secondary antibody.

    See Abreview

  • ab14705 staining MTCO1 - Mitochondrial Marker in rat cerebellum primary cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% TX-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/2000 in 1% goat serum, 0.1% TX100; PBS) for 16 hours at 4°C. An Alexa Fluor®546-conjugated Goat polyclonal to mouse IgG, dilution 1/5000, was used as secondary antibody.

    See Abreview

  • Overlay histogram showing HEK293 cells stained with ab14705 (red line). The cells were fixed with 80% methanol (5 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 (ab14705, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • All lanes : Anti-MTCO1 antibody [1D6E1A8] (ab14705)

    Lane 1 : Isolated mitochondria from Human heart at 5 µg
    Lane 2 : Isolated mitochondria from Bovine heart at 1 µg
    Lane 3 : Isolated mitochondria from Rat heart at 10 µg
    Lane 4 : Isolated mitochondria from Mouse heart at 5 µg

    Secondary
    Goat Anti-Mouse IgG

    Predicted band size : 57 kDa
    Observed band size : 40 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 70 kDa. We are unsure as to the identity of these extra bands.
  • ab14705 staining MTCO1 in skeletal muscle tissue by Immunohistochemistry (Frozen sections). Tissue sections were from from a patient with a single large deletion of the mtDNA and show a mosaic of complex IV positive and complex IV negative fibers.
  • ab14705 staining MTCO1 in Human colon tissue by Immunohistochemistry (Frozen sections). Tissue sections from a normal ageing patient show complex IV negative crypts due to clonal expansion of colonic stem cells bearing mutations in the mtDNA-encoded gene for complex IV.
  • IHC image of MTCO1 staining in human normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab14705, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ab14705 staining rat pancreas sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1% BSA for 10 minutes at 21°C, followed by staining with ab14705 at 1/1000 in TBS/BSA/azide for 2h at 21°C. A biotinylated goat anti-mouse polyclonal antibody at 1/200 was used as the secondary antibody. Positivity in exocrine glands appears to be intense at the cytplasm of adjacent cells. The cells of the Islet of Langerhan to the right have a diffuse, punctate positivity.

    See Abreview

References for Anti-MTCO1 antibody [1D6E1A8] (ab14705)

This product has been referenced in:
  • Davoudi M  et al. Complex I function and supercomplex formation are preserved in liver mitochondria despite progressive complex III deficiency. PLoS One 9:e86767 (2014). WB ; Mouse . Read more (PubMed: 24466228) »
  • Crooks DR  et al. Elevated FGF21 secretion, PGC-1a and ketogenic enzyme expression are hallmarks of iron-sulfur cluster depletion in human skeletal muscle. Hum Mol Genet 23:24-39 (2014). Human . Read more (PubMed: 23943793) »

See all 95 Publications for this product

Product Wall

Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Sample Mouse Cultured Cells (Mouse Embryonic Fibroblasts)
Specification Mouse Embryonic Fibroblasts
Permeabilization Yes - 0.5% Triton-X
Fixative Paraformaldehyde
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Verified customer

Submitted May 26 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% GEL)
Sample Human Cell lysate - whole cell (H460)
Specification H460
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
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Submitted May 26 2014

Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Sample Human Cultured Cells (H460)
Specification H460
Permeabilization Yes - 0.5% Triton-X
Fixative Paraformaldehyde
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Submitted May 26 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% GEL)
Sample Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblasts)
Specification Mouse Embryonic Fibroblasts
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
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Submitted May 26 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Chinese Hamster Cell (Fibroblasts)
Specification Fibroblasts
Permeabilization Yes - 0.1% TX-100
Fixative Formaldehyde
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Dr. Roman Lyakhovetsky

Verified customer

Submitted Dec 17 2013

Application Flow Cytometry
Fixation Beckman Coulter PerFix-nc
Permeabilization Yes - Beckman Coulter PerFix-nc
Sample Rhesus monkey Cell (MSC)
Specification MSC
Gating Strategy Bone marrow MSCs
Preparation Cell harvesting/tissue preparation method: Isolated and expanded MSCs
Sample buffer: MEM + 20% FBS
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Submitted Oct 21 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing
Sample Fruit fly (Drosophila melanogaster) Tissue lysate - whole (Head)
Specification Head
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Aug 09 2013

I have a few follow up questions regarding your protocol and troubleshooting tips:

1. The primary antibody contains sodium azide. Sodium azide inhibits HRP, the conjugate of your secondary antibody. Thus, you will need to either purify the ant...

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Application Western blot
Sample Mouse Tissue lysate - whole (DRG)
Loading amount 10 µg
Specification DRG
Gel Running Conditions Reduced Denaturing (12)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Jan 11 2013



I am sorry to hear that you have been experiencing issues when using this antibody. This product is covered by our Abpromise guarantee. According to Abpromise guarantee, if this product has been purchased within the last six months and is not...

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