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

Overview

  • Product nameAnti-VDAC1 / Porin antibody - Mitochondrial Loading ControlSee all VDAC1 / Porin primary antibodies ...
  • Description
    Rabbit polyclonal to VDAC1 / Porin - Mitochondrial Loading Control
  • SpecificityThis antibody detects all 3 isoforms of VDAC.
  • Tested applicationsWB, 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).Can be blocked with Human VDAC1 / Porin peptide (ab16131).
IHC-P Use at an assay dependent dilution.
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
    • 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 - Mitochondrial Loading Control images

  • 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).

  • 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 : Heart (Mouse) Tissue Lysate
    Lane 2 : Kidney (Mouse) Tissue Lysate
    Lane 3 : Skeletal Muscle (Mouse) Tissue Lysate (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:
  • Shephard F  et al. A mitochondrial location for haemoglobins--dynamic distribution in ageing and Parkinson's disease. Mitochondrion 14:64-72 (2014). WB ; Mouse . Read more (PubMed: 24333691) »
  • De Ita-Pérez D  et al. Synchronization by food access modifies the daily variations in expression and activity of liver GABA transaminase. Biomed Res Int 2014:590581 (2014). WB ; Rat . Read more (PubMed: 24809054) »

See all 35 Publications for this product

Product Wall

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

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 (colone)
Specification colone
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

ilan sela

Verified customer

Submitted Apr 19 2012

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 (Thyroid)
Specification Thyroid
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 100mM sodium citrate
Permeabilization No
Blocking step 10% NGS, 5%BSA in PBS as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Ms. Hadas Pahima

Verified customer

Submitted Apr 16 2012

Thank you for your call today and for your enquiry.

As we discussed, I would recommend either ab14744 or ab15895 for immunostaining of mitochondria in mouse cells-

http://www.abcam.com/COX-IV-antibody-20E8C12-Mitochondrial-Loading-C...

Read More

After looking at these antibodies I think that they will most likely be appropriate for your use; however it does depend on the type of experiment you are performing. Both of these antibodies are tested and guaranteed to work in immunocytochemistry, im...

Read More
Application Western blot
Sample Human Cell lysate - other (human dermal fibroblasts)
Loading amount 20 µg
Specification human dermal fibroblasts
Gel Running Conditions Reduced Denaturing (4-12% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Sep 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Forebrain)
Specification Forebrain
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Username

Dr. Markus Kipp

Verified customer

Submitted Aug 04 2011

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