Anti-VDAC1 / Porin antibody [20B12AF2] - Mitochondrial Loading Control (ab14734)

Overview

• Product nameAnti-VDAC1 / Porin antibody [20B12AF2] - Mitochondrial Loading ControlSee all VDAC1 / Porin primary antibodies ...
• Description
  Mouse monoclonal [20B12AF2] to VDAC1 / Porin - Mitochondrial Loading Control
• Tested applicationsWB, ICC, IHC-P, ICC/IF, Flow Cyt more details
• Species reactivity
  Reacts with: Mouse, Rat, Human, Fruit fly (Drosophila melanogaster)
  
• Immunogen

  Recombinant full length protein (Human)

Properties

• FormLiquid
• Storage instructionsStore at +4°C. Do not freeze.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: HEPES buffered saline
• Concentration information loading...
• PurityIgG fraction
• Purification notesNear 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 number20B12AF2
• IsotypeIgG2b
• Light chain typekappa
• Research Areas
  • Metabolism
  • Types of disease
  • Cancer

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

  • Signal Transduction
  • Metabolism
  • Mitochondrial

  • Isotype/Loading Controls
  • Loading Controls
  • VDAC1 - Mitochondrial Preps

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

Applications

Target

• FunctionForms a channel through the mitochondrial outer membrane and also the plasma membrane. The channel at the outer mitochondrial membrane allows diffusion of small hydrophilic molecules; in the plasma membrane it is involved in cell volume regulation and apoptosis. It adopts an open conformation at low or zero membrane potential and a closed conformation at potentials above 30-40 mV. The open state has a weak anion selectivity whereas the closed state is cation-selective. May participate in the formation of the permeability transition pore complex (PTPC) responsible for the release of mitochondrial products that triggers apoptosis.
• Tissue specificityHeart, liver and skeletal muscle.
• Sequence similaritiesBelongs to the eukaryotic mitochondrial porin family.
• DomainConsists mainly of a membrane-spanning beta-barrel formed by 19 beta-strands. The helical N-terminus folds back into the pore opening and plays a role in voltage-gated channel activity.
• Cellular localizationMitochondrion outer membrane. Cell membrane.

Target information above from: UniProt accession P21796 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 7416 Human
  • Entrez Gene: 22333 Mouse
  • Entrez Gene: 83529 Rat
  • Omim: 604492 Human
  • SwissProt: P21796 Human
  • SwissProt: Q60932 Mouse
  • SwissProt: Q9Z2L0 Rat
  • Unigene: 519320 Human

  • Unigene: 709372 Human
  • Unigene: 3555 Mouse
  • Unigene: 470023 Mouse
  • Unigene: 54594 Rat

  see all

• Alternative names
  hVDAC1 antibodyMGC111064 antibodyMitochondrial Porin antibody

  Outer mitochondrial membrane protein porin 1 antibodyPlasmalemmal porin antibodyPorin 31HL antibodyPorin 31HM antibodyPorin antibodyPORIN-31-HL antibodyVDAC 1 antibodyVDAC antibodyVDAC-1 antibodyVdac1 antibodyVDAC1_HUMAN antibodyVoltage Dependent Anion Channel 1 antibodyVoltage dependent anion selective channel protein 1 antibodyVoltage-dependent anion-selective channel protein 1 antibody

  see all

Anti-VDAC1 / Porin antibody [20B12AF2] - Mitochondrial Loading Control images

• 

  Immunocytochemistry/ Immunofluorescence - VDAC1 / Porin antibody [20B12] - Mitochondrial Loading Control (ab14734)

  Immunofluoresence using ab14734 at 0.2 µg/ml on human fibroblasts (red).
  Nuclei were labelled with DAPI (blue).

• 

  Western blot - VDAC1 / Porin antibody [20B12] - Mitochondrial Loading Control (ab14734)
  All lanes : Anti-VDAC1 / Porin antibody [20B12AF2] - Mitochondrial Loading Control (ab14734) at 1/5000 dilution
  
  Lane 1 : Rat brain cell lysate (homogenate)
  Lane 2 : Rat brain cell lysate (homogenate)
  Lane 3 : Rat brain cell lysate (mitochondrial)
  Lane 4 : Rat brain cell lysate (mitochondrial)
  
  Lysates/proteins at 30 µg per lane.
  
  Secondary
  HRP conjugated sheep anti-mouse IgG
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Observed band size : 39 kDa (why is the actual band size different from the predicted?)
  
  
  Exposure time : 5 minutes

  This image is courtesy of an anonymous Abreview

  See Abreview

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - VDAC1 / Porin antibody [20B12] - Mitochondrial Loading Control (ab14734)
  Ab14734 staining Human normal left ventricle. Staining is localized to the cytoplasm.
  Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
  Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
• 

  Flow Cytometry - VDAC1 / Porin antibody [20B12] - Mitochondrial Loading Control (ab14734)
  Overlay histogram showing HepG2 cells stained with ab14734 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab14734, 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 IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
  
  
• 

  Western blot - VDAC1 / Porin antibody [20B12] - Mitochondrial Loading Control (ab14734)
  All lanes : Anti-VDAC1 / Porin antibody [20B12AF2] - Mitochondrial Loading Control (ab14734)
  
  Lane 1 : Isolated mitochondria from human heart at 15 µg
  Lane 2 : Isolated mitochondria from bovine heart at 6 µg
  Lane 3 : Isolated mitochondria from rat heart at 30 µg
  Lane 4 : Isolated mitochondria from mouse heart at 30 µg
  Lane 5 : HepG2 at 30 µg
  
  
  Observed band size : 37 kDa (why is the actual band size different from the predicted?)