Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895)

Overview

  • Product nameAnti-VDAC1 / Porin antibody - Mitochondrial Loading Control
    See all VDAC1 / Porin primary antibodies
  • Description
    Rabbit polyclonal to VDAC1 / Porin - Mitochondrial Loading Control
  • SpecificityThis antibody detects all 3 isoforms of VDAC.
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Dog, Human, Zebrafish, Chinese Hamster
    Predicted to work with: Rabbit, Cow, Pig
  • Immunogen

    Synthetic peptide derived from residues 150 - 250 of Human VDAC1/ Porin. The immunogen sequence is completely conserved between isoforms 1, 2 and 3 of human VDAC (Note: the amino acid sequence is proprietary; Peptide available as ab16131.)

  • Positive control
    • This antibody gave a positive control in the following human whole cell lysates: HeLa A431 Jurkat HEK293 This antibody gave a positive control in HeLa (Human epithelial carcinoma cell line) Nuclear This antibody gave a positive control in the following mouse tissue lysates: Heart Kidney Skeletal Muscle Spinal Cord This antibody gave a positive control in the following rat lysates: PC12 Whole Cell Brain Tissue Lysate - normal tissue Kidney Whole Cell Lysate - normal tissue

Properties

Applications

Our Abpromise guarantee covers the use of ab15895 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 31 kDa (predicted molecular weight: 31 kDa).

Abcam recommends using BSA as the blocking agent.

IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.

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 - Mitochondrial Loading Control images

  • Image courtesy of Human Protein Atlas

    Paraffin embedded sections of human kidney were incubated with ab15895 (1/600 dilution) at room temperature for 30 mins. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab15895 was also tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further images can be found at www.proteinatlas.org

  • All lanes : Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895) at 1 µg/ml

    Lane 1 : Hela Nuclear lysate
    Lane 2 : Hela cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : Jurkat cell lysate
    Lane 5 : HEK293 cell lysate
    Lane 6 : Hela Nuclear lysate with Human VDAC1 / Porin peptide (ab16131) at 1 µg/ml
    Lane 7 : Hela cell lysate with Human VDAC1 / Porin peptide (ab16131) at 1 µg/ml
    Lane 8 : A431 cell lysate with Human VDAC1 / Porin peptide (ab16131) at 1 µg/ml
    Lane 9 : Jurkat cell lysate with Human VDAC1 / Porin peptide (ab16131) at 1 µg/ml
    Lane 10 : HEK293 cell lysate with Human VDAC1 / Porin peptide (ab16131) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 31 kDa
    Observed band size : 31 kDa
  • ICC/IF image of ab15895 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab15895, 5µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 phalloidin was used to label F-actin (red).

  • ab15895 staining VDAC1/Porin in Human Testis 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 citric acid. Samples were incubated with primary antibody (1/50 in TBS/BSA/azide buffer) for 16 hours at 21°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895) at 1 µg/ml

    Lane 1 : Heart (Mouse) Tissue Lysate
    Lane 2 : Kidney (Mouse) Tissue Lysate
    Lane 3 : Mouse skeletal muscle tissue lysate - total protein (ab29711)
    Lane 4 : Spinal Cord (Mouse) Tissue Lysate
    Lane 5 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 6 : Brain (Rat) Tissue Lysate - normal tissue
    Lane 7 : Kidney (Rat) Whole Cell Lysate - normal tissue (ab29480)

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 31 kDa
    Observed band size : 31 kDa
  • ab15895 staining mouse heart tissue section by IHC-P.  Sections were PFA fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking in 10% serum for 10 minutes at 25°C.  The primary antibody was diluted 1/600 and incubated with the sample for 1 hour at 25°C.  A biotinylated goat anti-rabbit antibody diluted 1/400 was used as the secondary.

    See Abreview

  • All lanes : Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895) at 1 µg/ml

    Lane 1 : Adult wild type zebrafish brain mitochondrial lysate at 20 µg
    Lane 2 : Adult wild type zebrafish brain cytosolic lysate at 30 µg

    Secondary
    HRP-conjugated goat F(ab’)2 Fragment at 1/10000 dilution
    developed using the ECL technique

    Predicted band size : 31 kDa
    Observed band size : 28,29 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    Image courtesy of an anonymous Abreview.

    See Abreview

  • ab15895 staining VDAC1 / Porin in human cerebellum tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in paraformaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then incubated with ab15895 at a 1/1000 dilution for 12 hours at 4°C. The secondary used was an undiluted HRP conjugated anti-rabbit polyclonal. Note: The big cells are purkinje cells of the cerebellum, (no nuclear staining).

    See Abreview


  • developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 31 kDa


    Exposure time : 3 minutes

    Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) (ab119220) with the
    Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623) .
    20ug of Lysate per lane and detection using ab15895 diluted to 1ug/ml.
    Lane 1: HeLa cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: A431 cell lysate
    Lane 4: HEK293 cell lysate
    Lane 5: HepG2 cell lysate.


  • developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 31 kDa

References for Anti-VDAC1 / Porin antibody - Mitochondrial Loading Control (ab15895)

This product has been referenced in:
  • Qiu W  et al. ATP Binding and Hydrolysis Properties of ABCB10 and Their Regulation by Glutathione. PLoS One 10:e0129772 (2015). Read more (PubMed: 26053025) »
  • Hou Y  et al. Rab27A Is Present in Mouse Pancreatic Acinar Cells and Is Required for Digestive Enzyme Secretion. PLoS One 10:e0125596 (2015). Mouse . Read more (PubMed: 25951179) »

See all 44 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - other (Hippocampus)
Gel Running Conditions Reduced Denaturing (13%)
Loading amount 6 µg
Specification Hippocampus
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Sergi Bayod

Verified customer

Submitted Aug 31 2016

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Zebrafish Tissue sections (heart)
Antigen retrieval step None
Permeabilization Yes - PBS+0.1% TWEEN
Specification heart
Blocking step Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted May 23 2016

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 (Testis)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Testis
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

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (e13 skin)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification e13 skin
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

Application Western blot
Sample Spermophilus tridecemlineatus Tissue lysate - other (Liver tissue fractions)
Gel Running Conditions Reduced Denaturing (10%)
Loading amount 20 µg
Specification Liver tissue fractions
Blocking step BSA as blocking agent for 8 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Username

Mr. James Bjork

Verified customer

Submitted Nov 03 2015

Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Denaturing (12 % acrylamide)
Sample Guinea pig Cell lysate - whole cell (Liver, cell lysate mitochondria)
Specification Liver, cell lysate mitochondria
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 7% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Jan 21 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Sample Mouse Tissue sections (Brain)
Specification Brain
Permeabilization Yes - 0.3% Triton X-100
Fixative Paraformaldehyde
Username

Prof. Chichung Lie

Verified customer

Submitted Aug 01 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 (Spleen)
Specification Spleen
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: CC1 Standard
Blocking step S Block as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: 37°C
Username

Abcam user community

Verified customer

Submitted Dec 11 2012

Thank you for bringing this to our attention. As I mentioned in our conversation, what you are seeing my be a doublet representing phosphorylated porin (upper band) and non-phosphorylated porin (lower band).

In any case, I have issued a free...

Read More
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 (Liver)
Specification Liver
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 100mM sodium citrate
Permeabilization No
Blocking step (agent) for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Ms. Angela Smilansy

Verified customer

Submitted Apr 19 2012

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