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

Overview

  • Product nameAnti-VDAC1 / Porin antibody [20B12AF2]
    See all VDAC1 / Porin primary antibodies
  • Description
    Mouse monoclonal [20B12AF2] to VDAC1 / Porin
  • Tested applicationsSuitable for: WB, ICC, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Goat, Cat, Dog, Human, Pig, Drosophila melanogaster, Fish, Quail, Marmoset (common), Dogfish/Catshark
  • Immunogen

    Recombinant full length protein (Human)

  • General notes

    Product was previously marketed under the MitoSciences sub-brand.

Properties

Applications

Our Abpromise guarantee covers the use of ab14734 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 39 kDa.
ICC Use a concentration of 0.2 µg/ml.
IHC-P Use a concentration of 2 µg/ml.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

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.
  • Information by UniProt
  • Database links
  • Alternative names
    • hVDAC1 antibody
    • Outer mitochondrial membrane protein porin 1 antibody
    • Plasmalemmal porin antibody
    • Porin 31HL antibody
    • Porin 31HM antibody
    • Porin antibody
    • VDAC 1 antibody
    • VDAC antibody
    • VDAC-1 antibody
    • Vdac1 antibody
    • VDAC1_HUMAN antibody
    • Voltage Dependent Anion Channel 1 antibody
    • Voltage dependent anion selective channel protein 1 antibody
    • Voltage-dependent anion-selective channel protein 1 antibody
    see all

Anti-VDAC1 / Porin antibody [20B12AF2] images

  • ab14734 staining human kidney sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/4000 in TBS/BSA/azide) for 2 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

  • ab14734 staining quail retina tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/2000 in TBS/BSA/azide) for 2 hours at 21°C. A Biotin-conjugated goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • ab14734 staining VDAC1 / Porin in human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 for 3 minutes and blocked with 0.2% serum for 60 minutes at 22°C. Samples were incubated with primary antibody (1/200 in 0.5% BSA and 0.02% TritonX100 in PBS) for 16 hours at 4°C. A FITC-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody at a dilution of 1/200.

  • All lanes : Anti-VDAC1 / Porin antibody [20B12AF2] (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?)
    Extra bands in the mouse sample (lane 4) are due to the reaction of the secondary antibody with residual mouse blood in the heart tissue, as it is very difficult to entirely remove the blood from these small organs.
  • 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.

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

  • Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734) at 1 µg/ml + Bovine Heart Mitochondria at 10 µg

    Secondary
    Goat anti-mouse-HRP (1:5000)
    Developed using the ECL technique

    Performed under reducing conditions.

  • Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 3 minutes

    Image courtesy of an anonymous abreview.

    Western blot analysis of human MCF-7 cell lysate (10μg/lane) labelling VDAC1 / Porin with ab14734 at 1/500 in 0.5% TBS-tween + Lait 5% NaN3 for 16 hours at 4ºC. A HRP-conjugated goat anti-mouse polyclonal (1/5000) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-VDAC1 / Porin antibody [20B12AF2] (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

References for Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)

This product has been referenced in:
  • Guo X  et al. VCP recruitment to mitochondria causes mitophagy impairment and neurodegeneration in models of Huntington's disease. Nat Commun 7:12646 (2016). Read more (PubMed: 27561680) »
  • Sen A & Cox RT Clueless is a conserved ribonucleoprotein that binds the ribosome at the mitochondrial outer membrane. Biol Open 5:195-203 (2016). Read more (PubMed: 26834020) »

See all 93 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Quail Tissue sections (Retina)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Retina
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted May 16 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Permeabilization Yes - 0.1% TritonX100 for 3mins
Specification HeLa
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 22°C
Fixative Paraformaldehyde
Username

Michiel Krols

Verified customer

Submitted Mar 02 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dogfish/Catshark Tissue sections (Intestine)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Intestine
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 29 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Goat Tissue sections (Myocardium)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Myocardium
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 29 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Marmoset (common) Tissue sections (Jejunum)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Jejunum
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 29 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (Myocardium)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Myocardium
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 29 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Fish Tissue sections (Skeletal muscle)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Skeletal muscle
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 12 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Gecko Tissue sections (Spinal cord)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Spinal cord
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 12 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Pig Tissue sections (Smooth muscle)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citric acid
Permeabilization No
Specification Smooth muscle
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Jan 28 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Sheep Tissue sections (Myocardium)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Myocardium
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Jan 28 2016

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