製品の概要

  • 製品名
    Anti-MTCO1 antibody [1D6E1A8]
    MTCO1 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [1D6E1A8] to MTCO1
  • 特異性
    In mouse liver lysate a specific band below 37 kDa was detected.
  • アプリケーション
    適用あり: ICC/IF, IHC-FoFr, IHC-P, WB, ICC, Flow Cyt, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Rat, Goat, Cow, Human, Pig, Caenorhabditis elegans, Zebrafish, Quail, Rhesus monkey, Chinese hamster
  • 免疫原

    Purified mitochondrial Complex IV subunit I (Human).

  • ポジティブ・コントロール
    • 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.
  • 特記事項

    Western blot protocol advice:
    For best results with this antibody in Western blot, do not boil samples before loading onto the gel. Boiling of the sample will cause a loss of signal.

    This antibody clone [1D6E1A8] is manufactured by Abcam.
    We have the following conjugates available:
    Anti-MTCO1 antibody (Alexa Fluor® 488) [1D6E1A8] (ab154477)
    Anti-MTCO1 antibody (Alexa Fluor® 647) [1D6E1A8] (ab198600)

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.

ターゲット情報

  • 機能
    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. 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.
  • パスウェイ
    Energy metabolism; oxidative phosphorylation.
  • 関連疾患
    Defects 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].
  • 配列類似性
    Belongs to the heme-copper respiratory oxidase family.
  • 細胞内局在
    Mitochondrion inner membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • COI antibody
    • COX I antibody
    • COX1 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

画像

  • ab14705 staining MTCO1 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 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

  • 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

  • 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.Extra bands in the mouse sample (lane 4) are due to the reaction of the IgG-specific goat anti-mouse secondary antibody with residual mouse blood in the heart tissue, as it is very difficult to entirely remove the blood from these small organs.
  • 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.

  • ab14705 staining MTCO1 in pig smooth muscle tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/250 in TBS/BSA/azide) for 2 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

    The image shows the two smooth muscle layers (inner circular is above the outer longitudinal) of the small intestine. Between the two layers one can see Ganglion cells of Aurbach's plexus. Both muscle and ganglion cells are enriched with mitochondria, as expected.

    See Abreview

参考文献

This product has been referenced in:
  • Shin JM  et al. Targeted deletion of Crif1 in mouse epidermis impairs skin homeostasis and hair morphogenesis. Sci Rep 7:44828 (2017). IHC-P ; Mouse . Read more (PubMed: 28317864) »
  • Huhta H  et al. Intratumoral lactate metabolism in Barrett's esophagus and adenocarcinoma. Oncotarget 8:22894-22902 (2017). IHC ; Human . Read more (PubMed: 28206968) »

See all 168 Publications for this product

レビューと Q&A

Application
Western blot
Sample
Drosophila melanogaster Tissue lysate - nuclear (brain)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (NuPage Gels 12% BT gels)
Loading amount
20 µg
Specification
brain
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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投稿 Sep 21 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Drosophila melanogaster Cell lysate - other (whole fly mitochondrial fraction)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
50 µg
Specification
whole fly mitochondrial fraction
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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投稿 Sep 12 2017

Application
Western blot
Sample
Mouse Cell lysate - other (Fibroblasts, mitochondrial fraction.)
Gel Running Conditions
Reduced Denaturing (18)
Loading amount
25 µg
Specification
Fibroblasts, mitochondrial fraction.
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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投稿 Sep 06 2017

Application
Western blot
Sample
American Robin (Turdis migratorius) Cell lysate - other (Fibroblasts, mitochondrial fraction.)
Gel Running Conditions
Reduced Denaturing (18)
Loading amount
25 µg
Specification
Fibroblasts, mitochondrial fraction.
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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投稿 Sep 06 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Retina)
Permeabilization
Yes - 0.3% Triton
Specification
Retina
Blocking step
5% Donkey Serum, 2% BSA, 0.3% Triton as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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投稿 Aug 09 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Retina, RPE)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Retina, RPE
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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投稿 Jul 25 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse retina)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
Mouse retina
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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投稿 Oct 18 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (ARPE19, hfRPE)
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
20 µg
Specification
ARPE19, hfRPE
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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投稿 Oct 18 2016

Application
Western blot
Sample
Zebrafish Tissue lysate - whole (Retina)
Gel Running Conditions
Reduced Denaturing (10%, hand-cast)
Loading amount
40 µg
Treatment
M-PER for 30 minutes
Specification
Retina
Blocking step
Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C
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投稿 Jun 03 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Goat Tissue sections (Heart muscle)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Heart muscle
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Feb 29 2016

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

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