Anti-COX IV antibody [20E8C12] - Mitochondrial Loading Control (ab14744)

Overview

• Product nameAnti-COX IV antibody [20E8C12] - Mitochondrial Loading ControlSee all COX IV primary antibodies ...
• Description
  Mouse monoclonal [20E8C12] to COX IV - Mitochondrial Loading Control
• Tested applicationsFlow Cyt, IHC-FoFr, WB, ICC/IF, IHC-Fr, IHC-P, IP more details
• Species reactivity
  Reacts with: Mouse, Rat, Hamster, Cow, Human, Zebrafish
  
• Immunogen

  Purified mitochondrial Complex IV (Cow).

• Positive controlHuman, bovine, murine and rat heart mitochondria.
• General notes

  
  

  This antibody makes an effective loading control for mitochondria. COXIV is generally expressed at a consistent high level. However, be aware that many proteins run at the same 16kD size as COXIV - 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 Rabbit polyclonal to COX IV antibody (ab16056).

Properties

• FormLiquid
• Storage instructionsStore at +4°C.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: HEPES
• Concentration information loading...
• PurityIgG fraction
• Primary antibody notes This antibody makes an effective loading control for mitochondria. COXIV is generally expressed at a consistent high level. However, be aware that many proteins run at the same 16kD size as COXIV - 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 Rabbit polyclonal to COX IV antibody (ab16056).
• Clonality Monoclonal
• Clone number20E8C12
• IsotypeIgG2a
• Light chain typekappa
• Research Areas
  • Metabolism
  • Types of disease
  • Cancer

  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Oxidative phosphorylation
  • Complex IV

  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Mitochondrial markers

  • Metabolism
  • Pathways and Processes
  • Metabolic signaling pathways
  • Energy transfer pathways
  • Integration of energy

  • Cancer
  • Cancer Metabolism
  • Metabolic signaling pathway
  • Integration of energy metabolism

  • Isotype/Loading Controls
  • Loading Controls
  • COX IV - Mitochondrial

  • Tags & Cell Markers
  • Subcellular Markers
  • Organelles
  • Mitochondria

Applications

Target

• FunctionThis protein is one of the nuclear-coded polypeptide chains of cytochrome c oxidase, the terminal oxidase in mitochondrial electron transport.
• Tissue specificityUbiquitous.
• Sequence similaritiesBelongs to the cytochrome c oxidase IV family.
• Cellular localizationMitochondrion inner membrane.

Target information above from: UniProt accession P13073 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 281090 Cow
  • Entrez Gene: 1327 Human
  • Entrez Gene: 12857 Mouse
  • Entrez Gene: 29445 Rat
  • Omim: 123864 Human
  • SwissProt: P00423 Cow
  • SwissProt: Q3SZS0 Cow
  • SwissProt: P13073 Human

  • SwissProt: P19783 Mouse
  • SwissProt: P10888 Rat
  • Unigene: 433419 Human
  • Unigene: 386758 Mouse
  • Unigene: 2528 Rat

  see all

• Alternative names
  COX 4 antibodyCOX 4I1 antibodyCOX 4I2 antibody

  COX IV 1 antibodyCOX IV-1 antibodyCOX4 2 antibodyCOX4 antibodyCOX41_HUMAN antibodyCOX4B antibodyCOX4I1 antibodyCOX4I2 antibodyCOX4L2 antibodyCOXIV 2 antibodyCOXIV antibodyCytochrome c oxidase polypeptide IV antibodyCytochrome c oxidase subunit 4 isoform 1 antibodyCytochrome c oxidase subunit 4 isoform 1 mitochondrial antibodyCytochrome C Oxidase Subunit IV antibodyCytochrome c oxidase subunit IV isoform 1 antibodyCytochrome c oxidase subunit IV isoform 2 (lung) antibodydJ857M17.2 antibodyMGC72016 antibodymitochondrial antibody

  see all

Anti-COX IV antibody [20E8C12] - Mitochondrial Loading Control images

• 

  Flow Cytometry - COX IV antibody [20E8] - Mitochondrial Loading Control (ab14744)
  Overlay histogram showing HeLa cells stained with ab14744 (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 (ab14744, 0.5µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  
  
• 

  Western blot - COX IV antibody [20E8] - Mitochondrial Loading Control (ab14744)
  All lanes : Anti-COX IV antibody [20E8C12] - Mitochondrial Loading Control (ab14744) at 1/5000 dilution
  
  Lane 1 : Cytoplasmic fraction mouse NIH/3T3 cell lysate
  Lane 2 : Nuclear fraction mouse NIH/3T3 cell lysate
  
  Secondary
  HRP conjugated goat anti-mouse antibody
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Observed band size : 16 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 3 minutes

  See Abreview

• 

  Immunocytochemistry/ Immunofluorescence - COX IV antibody [20E8] - Mitochondrial Loading Control (ab14744)This image is courtesy of an anonymous Abreview

  ab14744 at 1/100 staining MCF-10A cells (human mammary epithelial cell line) by ICC/IF. The cells were methanol/acetone fixed at -20C for 5 minutes, blocked with BSA and then incubated with the antibody for 16 hours. The positive tissue was colocalised with a mitotracker (mitotrackers are a series of patented mt-selective stains that are concentrated by active mt and well retained during cell fixation). An Alexa-Fluor ® 488 conjugated goat anti-mouse antibody was used as the secondary. The image shows COXIV staining in green and DAPI staining in blue.

  See Abreview

• 

  Immunohistochemistry (PFA perfusion fixed frozen sections) - COX IV antibody [20E8] - Mitochondrial Loading Control (ab14744)Image from Hashimoto T. et. al., PLoS ONE. 2008; 3(8): e2915 (Fig. 1B)

  ab14744 staining COX IV in rat brain tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Rats were anesthetized and intracardially perfused with 500 ml of normal saline at room temperature, followed by 500 ml of ice-cold, freshly made 4% paraformaldehyde in phosphate buffer (PB, 0.1 M, pH 7.4). A Cy3 conjugated anti mouse antibody was used as secondary. 

• 

  Western blot - COX IV antibody [20E8] - Mitochondrial Loading Control (ab14744)
  All lanes : Anti-COX IV antibody [20E8C12] - Mitochondrial Loading Control (ab14744) at 1 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 murine heart at 10 µg
  
  
  Observed band size : 16 kDa (why is the actual band size different from the predicted?)
  
• 

  Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [20E8C12] - Mitochondrial Loading Control (ab14744)This image is courtesy of an anonymous Abreview
  ab14744 staining COX IV - Mitochondrial Loading in the HEK293 cell line from Human embryonic kidney by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 1% in PBS and blocked with 5% serum for 60 minutes at 21°C. Samples were incubated with primary antibody (1/200 in 5% Goat serum) for 2 hours. An Alexa Fluor^®546-conjugated Goat anti-mouse IgG polyclonal was used as the secondary antibody.

  See Abreview