Anti-MTCO1 antibody [1D6E1A8] (ab14705)
![Immunocytochemistry - Anti-MTCO1 antibody [1D6E1A8] (ab14705)](https://www.abcam.com/ps/products/14/ab14705/Images/ab14705-13-anti-mtco1-1d6ee1a8-antibody-immunocytochemistry-hela-human.jpg "Immunocytochemistry - Anti-MTCO1 antibody [1D6E1A8] (ab14705)")
Key features and details
- Mouse monoclonal [1D6E1A8] to MTCO1
- Suitable for: ICC, IHC-P, WB, Flow Cyt
- Reacts with: Mouse, Rat, Cow, Human
- Isotype: IgG2a
Related conjugates and formulations
Overview
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Product name
Anti-MTCO1 antibody [1D6E1A8]
See all MTCO1 primary antibodies -
Description
Mouse monoclonal [1D6E1A8] to MTCO1 -
Host species
Mouse -
Specificity
In mouse liver lysate a specific band below 37 kDa was detected. -
Tested applications
Suitable for: ICC, IHC-P, WB, Flow Cytmore details -
Species reactivity
Reacts with: Mouse, Rat, Cow, Human
Predicted to work with: Sheep, Goat, Cat, Dog, Pig, Caenorhabditis elegans, Zebrafish, Quail, Rhesus monkey, Chinese hamster, Common marmoset
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Immunogen
Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human cerebellar and colon tissue. WB: Mouse, human, bovine and rat heart mitochondria lysate. Flow Cyt: HEK-293 cells. ICC: HeLa cells.
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General notes
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.Hydrophobic intrinsic membrane proteins such as the core mtDNA-encoded proteins of the mitochondrial OXPHOS complexes tend to run faster in SDS-PAGE than predicted by their amino acid composition. This is likely due to incomplete unfolding of the protein and a more negative charge:mass ratio.
This antibody clone [1D6E1A8] is manufactured by Abcam. If you require a different buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com.
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. -
Storage buffer
pH: 7.5
Preservative: 0.02% Sodium azide
Constituent: HEPES buffered saline -
Concentration information loading... -
Purity
Ammonium Sulphate Precipitation -
Purification notes
Near homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation. -
Clonality
Monoclonal -
Clone number
1D6E1A8 -
Isotype
IgG2a -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
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 | (2) |
Use a concentration of 1 µg/ml.
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| IHC-P | (13) |
Use a concentration of 5 µg/ml.
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| WB | (18) |
Use a concentration of 0.5 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 57 kDa).
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| Flow Cyt | (1) |
Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
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ICC
Use a concentration of 1 µg/ml. |
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IHC-P
Use a concentration of 5 µg/ml. |
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WB
Use a concentration of 0.5 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 57 kDa). |
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Flow Cyt
Use 1µg for 106 cells. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
Target
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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. 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. -
Pathway
Energy metabolism; oxidative phosphorylation. -
Involvement in disease
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]. -
Sequence similarities
Belongs to the heme-copper respiratory oxidase family. -
Cellular localization
Mitochondrion inner membrane. - Information by UniProt
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Database links
- Entrez Gene: 2565700 Caenorhabditis elegans
- Entrez Gene: 281919 Cow
- Entrez Gene: 4512 Human
- Entrez Gene: 17708 Mouse
- Entrez Gene: 26195 Rat
- Entrez Gene: 140539 Zebrafish
- Omim: 516030 Human
- SwissProt: P24893 Caenorhabditis elegans
see all -
Alternative names
- COI antibody
- COX I antibody
- COX1 antibody
see all
Images
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Immunocytochemistry - Anti-MTCO1 antibody [1D6E1A8] (ab14705)
ab14705 staining MTCO1 in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab14705 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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![Western blot - Anti-MTCO1 antibody [1D6E1A8] (ab14705)](https://www.abcam.com/ps/products/14/ab14705/Images/ab14705-22033-anti-mtco1-antibody-1d6e1a8-western-blot.jpg)
Western blot - Anti-MTCO1 antibody [1D6E1A8] (ab14705)All lanes : Anti-MTCO1 antibody [1D6E1A8] (ab14705) at 0.5 µg/ml
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
All lanes : 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. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTCO1 antibody [1D6E1A8] (ab14705)](https://www.abcam.com/ps/products/14/ab14705/Images/ab14705-22030-anti-mtco1-antibody-1d6e1a8-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTCO1 antibody [1D6E1A8] (ab14705)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.
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![Flow Cytometry - Anti-MTCO1 antibody [1D6E1A8] (ab14705)](https://www.abcam.com/ps/products/14/ab14705/Images/ab14705-22034-anti-mtco1-antibody-1d6e1a8-flow-cytometry.jpg)
Flow Cytometry - Anti-MTCO1 antibody [1D6E1A8] (ab14705)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. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTCO1 antibody [1D6E1A8] (ab14705)](https://www.abcam.com/ps/products/14/ab14705/Images/ab14705-353479-AntiMTCO1-antibody-1D6E1A8-immunohistochemistry-human.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTCO1 antibody [1D6E1A8] (ab14705)Image from Phillips J et al., Sci Rep. 2016 May 16;6:26013. Fig 3b doi: 10.1038/srep26013. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.250 μm human cerebellar sections from control individuals and a patient with mitochondrial disease underwent passive clearing at 37 °C for 2 or 4 weeks.
The quality of immunofluorescent staining is determined by duration of passive clearing; 2 weeks of passive clearing produced minimal labelling of the white matter in the granule cell layer (NF-H; green; 488 nm and MBP; red, 546 nm) with an absence of labelling of mitochondria (MTCO1 (COXI) (ab14705, 1/100); purple; 647 nm; Extending passive clearing to 4 weeks improved the quality of stain with identifiable Purkinje cells and their axons (NF-H, green; 488 nm) and their myelin sheaths (MBP; red, 546 nm) and mitochondria (MTCO1 (COXI) (ab14705, 1/100); purple; 647 nm.
Datasheets and documents
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SDS download
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Datasheet download
References (409)
ab14705 has been referenced in 409 publications.
- He Q et al. Mtu1 defects are correlated with reduced osteogenic differentiation. Cell Death Dis 12:61 (2021). PubMed: 33431792
- Iwasaki H et al. MicroRNA-494-3p inhibits formation of fast oxidative muscle fibres by targeting E1A-binding protein p300 in human-induced pluripotent stem cells. Sci Rep 11:1161 (2021). PubMed: 33441918
- Romani M et al. NAD+ boosting reduces age-associated amyloidosis and restores mitochondrial homeostasis in muscle. Cell Rep 34:108660 (2021). PubMed: 33472069
- Yagi M et al. Mitochondrial translation deficiency impairs NAD+ -mediated lysosomal acidification. EMBO J 40:e105268 (2021). PubMed: 33528041
- Cardoso S et al. IGF1R Deficiency Modulates Brain Signaling Pathways and Disturbs Mitochondria and Redox Homeostasis. Biomedicines 9:N/A (2021). PubMed: 33562061