MitoSciences (MSA03)

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 applicationsWB, ICC, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Fruit fly (Drosophila melanogaster)
  • Immunogen

    Recombinant full length protein (Human)

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.

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
    • MGC111064 antibody
    • Mitochondrial Porin antibody
    • Outer mitochondrial membrane protein porin 1 antibody
    • Plasmalemmal porin antibody
    • Porin 31HL antibody
    • Porin 31HM antibody
    • Porin antibody
    • PORIN-31-HL 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

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

  • 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

  • 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.
  • 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.

  • 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?)

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

This product has been referenced in:
  • Viola HM  et al. Impaired functional communication between the L-type calcium channel and mitochondria contributes to metabolic inhibition in the mdx heart. Proc Natl Acad Sci U S A 111:E2905-14 (2014). IP ; Mouse . Read more (PubMed: 24969422) »
  • Yun J  et al. MUL1 acts in parallel to the PINK1/parkin pathway in regulating mitofusin and compensates for loss of PINK1/parkin. Elife (Cambridge) 3:e01958 (2014). WB ; Human, Fruit fly (Drosophila melanogaster) . Read more (PubMed: 24898855) »

See all 75 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Sample Mouse Tissue lysate - whole (Mouse brain)
Specification Mouse brain
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Jun 02 2014

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Sample Human Cell (hek293)
Specification hek293
Permeabilization Yes - 1% triton x100
Fixative Paraformaldehyde
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Submitted Oct 04 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
Sample Mouse Cell (MEF)
Specification MEF
Permeabilization Yes - 1%triton x100
Fixative Paraformaldehyde
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Submitted Oct 04 2013

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Human Cell lysate - whole cell (hek293)
Specification hek293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
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Submitted Oct 04 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Sample Rat Cell (H9C2)
Specification H9C2
Permeabilization Yes - 1% TRITON X100
Fixative Paraformaldehyde
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Submitted Oct 04 2013

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Cell lysate - whole cell (h9c2)
Specification h9c2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
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Submitted Oct 04 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing
Sample Human Tissue lysate - whole (Skeletal Muscle)
Specification Skeletal Muscle
Treatment 12% Bis-Tris gel
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
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Submitted Sep 27 2013



Yes it is a standard mouse monoclonal IgG type 2b with heavy & light chains, FAb and Fc portions.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (DRG)
Loading amount 10 µg
Specification DRG
Gel Running Conditions Reduced Denaturing (12)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Jan 08 2013

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 (Skeletal Muscle)
Specification Skeletal Muscle
Fixative Methacarn
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9.0
Permeabilization No
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 23°C
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Submitted Dec 21 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"