Overview

  • Product name
    Anti-VDAC1 / Porin antibody [20B12AF2]
    See all VDAC1 / Porin primary antibodies
  • Description
    Mouse monoclonal [20B12AF2] to VDAC1 / Porin
  • Host species
    Mouse
  • Tested applications
    Suitable 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, Common marmoset, Dogfish, Catshark
  • Immunogen

    Recombinant full length protein corresponding to Human VDAC1/ Porin.

  • Positive control
    • WB: Isolated mitochondria from human, cow, rat and mouse heart. HepG2 cell lysate. IHC-P: Human kidney tissue. ICC/IF: HeLa cells. Human fibroblasts. Flow Cyt: HepG2 cells.
  • General notes

    This antibody clone [20B12AF2] is manufactured by Abcam.

    If you require this antibody in a different buffer formulation or a different conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    Abcam recommended secondaries - Goat Anti-Mouse HRP (ab205719) and Goat Anti-Mouse Alexa Fluor® 488 (ab150113).

    See other anti-mouse secondary antibodies that can be used with this antibody.

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

  • Function
    Forms 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 specificity
    Heart, liver and skeletal muscle.
  • Sequence similarities
    Belongs to the eukaryotic mitochondrial porin family.
  • Domain
    Consists 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 localization
    Mitochondrion outer membrane. Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • hVDAC1 antibody
    • mVDAC5 antibody
    • OMP2 antibody
    • Outer mitochondrial membrane protein porin 1 antibody
    • Plasmalemmal porin antibody
    • Porin 31HL antibody
    • Porin 31HM antibody
    • Porin antibody
    • VDAC 1 antibody
    • VDAC 5 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

Images

  • Double immunofluorescent staining and immunoblot analysis of the endogenous SCPX and SCP2 proteins in MA-10 cells.

    Double immunofluorescent staining of endogenous SCPX in MA-10 cells (red), PMP70 (peroxisomal marker protein, painted in green), and VDAC1 (mitochondrial marker protein, painted in green, ab14734). The nucleus was counterstained with DAPI (painted in green).

    Scale bar, 5 μm.

  • Double immunofluorescent staining and immunoblot analysis of the endogenous SCPX and SCP2 proteins in MA-10 cells.

    Immunoblot analysis of endogenous SCP2 in subcellular fractions of MA-10 cell lysates. VDAC1 is used as a mitochondrial marker protein, and GAPDH is used as a whole cell lysate loading control.

    Lane 1: Whole cell lysate

    Lane 2: Cytosolic fraction

    Lane 3: Mitochondrial-enriched fraction

  • Quantification of the fiber type specific mitochondrial in mouse, rat and human skeletal muscles using different marker of mitochondrial content.

    In situ immunolabeling of a human vastus lateralis cross-section for MHC type IIx and laminin (green), type I (Blue) and type IIa (red) (M) and its corresponding VDAC (a marker of mitochondrial content) immunolabeling performed on a serial cross-section (N).

  • 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 (Human liver hepatocellular carcinoma cell line) cell lysate 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.
  • 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 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.

    See Abreview

  • ab14734 staining VDAC1 / Porin in human HeLa (Human epithelial cell line from cervix adenocarcinoma) 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% Triton X100 in PBS) for 16 hours at 4°C. An FITC-conjugated goat anti-mouse IgG polyclonal was used as the secondary antibody at a dilution of 1/200.

    See Abreview

  • Overlay histogram showing HepG2 (Human liver hepatocellular carcinoma cell line) 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, 1 µg/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, 2 µg/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.

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

  • Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734) at 1/500 dilution + MCF7 (Human breast adenocarcinoma cell line) cell lysate at 10 µg

    Secondary
    HRP-conjugated goat anti-mouse polyclonal at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 3 minutes


    0.5% TBS-tween + Lait 5% NaN3 for 16 hours at 4ºC.

    See Abreview

  • All lanes : Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734) at 1/5000 dilution

    Lanes 1-2 : Rat brain cell lysate (homogenate)
    Lanes 3-4 : Rat brain cell lysate (mitochondrial)

    Lysates/proteins at 30 µg per lane.

    Secondary
    All lanes : 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

    See Abreview

References

This product has been referenced in:
  • Vazquez Fonseca L  et al. Mutations in COQ8B (ADCK4) found in patients with steroid-resistant nephrotic syndrome alter COQ8B function. Hum Mutat 39:406-414 (2018). Read more (PubMed: 29194833) »
  • Velazquez-Villegas LA  et al. TGR5 signalling promotes mitochondrial fission and beige remodelling of white adipose tissue. Nat Commun 9:245 (2018). Read more (PubMed: 29339725) »
See all 229 Publications for this product

Customer reviews and Q&As

1-10 of 58 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (C20 human microglia cell line)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
15 µg
Specification
C20 human microglia cell line
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Oct 17 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse brain)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
mouse brain
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted Oct 17 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate ph 6
Permeabilization
No
Specification
Brain tissue
Blocking step
Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Fixative
Formaldehyde

Herr Dr. Markus Kipp

Verified customer

Submitted Sep 07 2018

Application
Western blot
Sample
Gerbil Cell lysate - whole cell (Gerbil mouse tissues (Malacothrix typical))
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
Gerbil mouse tissues (Malacothrix typical)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted May 24 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Dog Cell lysate - whole cell (MDCK2 cells)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
20 µg
Specification
MDCK2 cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Herr Dr. Vladimir Milenkovic

Verified customer

Submitted May 16 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Rat Cultured Cells (Cardiomyocytes)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 21 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (heart)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 01 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain tissue lysate)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (3-8% Bis-Tris)
Loading amount
20 µg
Specification
Whole brain tissue lysate
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Apr 19 2017

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

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

Michiel Krols

Verified customer

Submitted Mar 02 2016

1-10 of 58 Abreviews or Q&A

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