Anti-COX IV抗体- Mitochondrial Loading Control (ab16056)

概述

  • 产品名称Anti-COX IV抗体- Mitochondrial Loading Control
    参阅全部 COX IV 一抗
  • 描述
    兔多克隆抗体to COX IV - Mitochondrial Loading Control
  • 经测试应用适用于: ICC, WB, ICC/IF, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Xenopus laevis, Potato, African Green Monkey, Chinese Hamster
    预测可用于: Chimpanzee
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human COX IV.

    (Peptide available as ab16381.)

  • 阳性对照
    • This antibody gave a positive signal when tested against: (WB) HeLa and Human Skeletal Muscle Tissue; (IHC-P) Human Liver Tissue; (IF) HeLa.
  • 常规说明

    This antibody makes an effective loading control for mitochondria. COX IV is generally expressed at a consistent high level. However, be aware that many proteins run at the same 16kD size as COX IV - our VDAC1 / Porin antibody makes a good alternative mitochondrial loading control for proteins of this size. Some caution is required when using this antibody as a loading control as COXIV expression can vary under some manipulations. An alternative mitochondrial loading control is Mouse monoclonal to COX IV antibody [20E8] (ab14744).

性能

应用

Our Abpromise guarantee covers the use of ab16056 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC Use at an assay dependent concentration.
WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 17 kDa).Can be blocked with Human COX IV peptide (ab16381).
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

靶标

  • 功能This protein is one of the nuclear-coded polypeptide chains of cytochrome c oxidase, the terminal oxidase in mitochondrial electron transport.
  • 组织特异性Ubiquitous.
  • 序列相似性Belongs to the cytochrome c oxidase IV family.
  • 细胞定位Mitochondrion inner membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • AL024441 antibody
    • COX 4 antibody
    • COX IV 1 antibody
    • COX IV antibody
    • COX IV-1 antibody
    • Cox4 antibody
    • COX41_HUMAN antibody
    • Cox4a antibody
    • COX4B antibody
    • COX4I1 antibody
    • COX4I2 antibody
    • COX4L2 antibody
    • COXIV antibody
    • Cytochrome c oxidase polypeptide IV antibody
    • Cytochrome c oxidase subunit 4 isoform 1 mitochondrial antibody
    • Cytochrome c oxidase subunit 4 isoform 1, mitochondrial antibody
    • Cytochrome C Oxidase subunit IV antibody
    • Cytochrome c oxidase subunit IV isoform 1 antibody
    • Cytochrome c oxidase subunit IV isoform 2 (lung) antibody
    • Cytochrome c oxydase subunit 4 antibody
    • dJ857M17.2 antibody
    • MGC105470 antibody
    • MGC72016 antibody
    see all

Anti-COX IV antibody - Mitochondrial Loading Control 图像

  • ab16056 staining COX IV in Mouse heart tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/600 ) for 12 hours at 4°C. A Cy5® donkey anti-rabbit secondary antibody was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab16056 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab16056, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red).

  • All lanes : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human liver tissue lysate - total protein (ab29889)
    Lane 3 : Human heart tissue lysate - total protein (ab29431)
    Lane 4 : Human skeletal muscle tissue lysate - total protein (ab29330)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 17 kDa
  • ab16056 staining COX IV in Mouse heart tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 5% serum for 1 hour at 21°C. Samples were incubated with primary antibody (1/200) for 12 hours at 4°C. An Alexa Fluour® 594 donkey anti-rabbit secondary antibody was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-COX IV antibody - Mitochondrial Loading Control (ab16056) at 0.38 µg/ml

    Lane 1 : HeLa whole cell lysate
    Lane 2 : Human skeletal muscle cell lysate
    Lane 3 : HeLa whole cell lysate with Human COX IV peptide (ab16381) at 1 µg/ml
    Lane 4 : Human skeletal muscle cell lysate with Human COX IV peptide (ab16381) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/10000 dilution

    Predicted band size : 17 kDa
    Observed band size : 15 kDa (why is the actual band size different from the predicted?)
  • IHC image of COXIV staining in human liver FFPE section, performed on a 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 ab16056, 1µg/ml, for 8 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.
  • ab16056 staining COX IV in breast tumour tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% buffered formalin and blocked with 5% serum for 60 minutes at 21°C; antigen retrieval was by heat mediation in a 10mM sodium citrate buffer pH6. Samples were incubated with primary antibody (1/300 in blocking buffer) for 12 hours at 4°C. An Alexa Fluor® 647-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

Anti-COX IV antibody - Mitochondrial Loading Control (ab16056)参考文献

This product has been referenced in:
  • Martin GG  et al. Fabp-1 gene ablation impacts brain endocannabinoid system in male mice. J Neurochem N/A:N/A (2016). Read more (PubMed: 27167970) »
  • Martin GG  et al. Female Mice are Resistant to Fabp1 Gene Ablation-Induced Alterations in Brain Endocannabinoid Levels. Lipids N/A:N/A (2016). WB ; Mouse . Read more (PubMed: 27450559) »

See all 52 Publications for this product

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (carotid body)
Permeabilization Yes - 0.5% TritonX in PBS
Specification carotid body
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative 4% PFA
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Abcam user community

Verified customer

提交于 Oct 06 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Bone marrow)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM Citrate buffer pH6
Permeabilization No
Specification Bone marrow
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative 10% normal buffered formalin
Username

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Verified customer

提交于 Mar 08 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (breast tumor)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM Sodium citrate buffer pH6
Permeabilization No
Specification breast tumor
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative 10% buffered formalin
Username

Abcam user community

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提交于 Mar 07 2016

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (heart)
Permeabilization No
Specification heart
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative Acetone
Username

Abcam user community

Verified customer

提交于 Oct 29 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (heart)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer pH6
Permeabilization No
Specification heart
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative Paraformaldehyde
Username

Abcam user community

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提交于 Oct 02 2015

Application Immunocytochemistry
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Sample Mouse Cultured Cells (Cardiomyocytes)
Specification Cardiomyocytes
Permeabilization Yes - Triton x-100, 0.01%
Fixative Formaldehyde
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提交于 Jan 16 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (heart)
Specification heart
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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Abcam user community

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提交于 Jan 16 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Human Tissue lysate - whole (skeletal muscle)
Specification skeletal muscle
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Rafael Montes Nieto

Verified customer

提交于 Oct 31 2014

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (hek293)
Specification hek293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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提交于 Jun 23 2014

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