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

  • ab16056 stained in Hela cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab16056 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature

  • 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.

    See Abreview

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

This product has been referenced in:
  • Liu T  et al. Molecular and Cellular Mechanisms of Apoptosis during Dissociated Spermatogenesis. Front Physiol 8:188 (2017). WB . Read more (PubMed: 28424629) »
  • Wang H  et al. Embryonic lethal abnormal vision proteins and adenine and uridine-rich element mRNAs after global cerebral ischemia and reperfusion in the rat. J Cereb Blood Flow Metab 37:1494-1507 (2017). Rat . Read more (PubMed: 27381823) »

See all 94 Publications for this product

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Application
Western blot
Sample
Pig Cell lysate - other (Porcine kidney epithelial cells (PK-15))
Gel Running Conditions
Reduced Denaturing (4-20% Tris-Glycine)
Loading amount
10 µg
Treatment
100 μg/ml poly(I:C) for 18 hrs
Specification
Porcine kidney epithelial cells (PK-15)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Sam Hardy

Verified customer

提交于 Jan 16 2017

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
Username

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

Abcam user community

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

Verified customer

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

Verified customer

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
Username

Abcam user community

Verified customer

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%
Username

Abcam user community

Verified customer

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

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