Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) is a mouse monoclonal antibody detecting VDAC1/Porin in Western Blot, Flow Cytometry, ICC/IF. Suitable for Cow, Human, Mouse, Rat.
- KO validated for confirmed specificity
- Over 580 publications
- Trusted since 2005
pH: 7.5
Preservative: 0.02% Sodium azide
Constituents: 0.88% Sodium chloride, 0.36% HEPES
Flow Cyt | WB | ICC/IF | |
---|---|---|---|
Human | Tested | Tested | Tested |
Mouse | Expected | Tested | Expected |
Rat | Expected | Tested | Expected |
Cat | Predicted | Predicted | Predicted |
Catshark | Predicted | Predicted | Predicted |
Common marmoset | Predicted | Predicted | Predicted |
Cow | Expected | Tested | Expected |
Dog | Predicted | Predicted | Predicted |
Dogfish | Predicted | Predicted | Predicted |
Drosophila melanogaster | Predicted | Predicted | Predicted |
Fish | Predicted | Predicted | Predicted |
Goat | Predicted | Predicted | Predicted |
Pig | Predicted | Predicted | Predicted |
Quail | Predicted | Predicted | Predicted |
Sheep | Predicted | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg for 106 Cells | Notes ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Cow | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Sheep, Goat, Cat, Dog, Pig, Drosophila melanogaster, Fish, Quail, Common marmoset, Dogfish, Catshark | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1 µg/mL | Notes - |
Species Rat | Dilution info 1 µg/mL | Notes - |
Species Human | Dilution info 1 µg/mL | Notes - |
Species Cow | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Sheep, Goat, Cat, Dog, Pig, Drosophila melanogaster, Fish, Quail, Common marmoset, Dogfish, Catshark | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Cow | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Sheep, Goat, Cat, Dog, Pig, Drosophila melanogaster, Fish, Quail, Common marmoset, Dogfish, Catshark | Dilution info - | Notes - |
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Non-selective voltage-gated ion channel that mediates the transport of anions and cations through the mitochondrion outer membrane and plasma membrane (PubMed:10661876, PubMed:11845315, PubMed:18755977, PubMed:30061676, PubMed:8420959). 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 (PubMed:10661876, PubMed:11845315, PubMed:18755977, PubMed:8420959). It adopts an open conformation at low or zero membrane potential and a closed conformation at potentials above 30-40 mV (PubMed:10661876, PubMed:18755977, PubMed:8420959). The open state has a weak anion selectivity whereas the closed state is cation-selective (PubMed:18755977, PubMed:8420959). Binds various signaling molecules, including the sphingolipid ceramide, the phospholipid phosphatidylcholine, and the sterols cholesterol and oxysterol (PubMed:18755977, PubMed:31015432). In depolarized mitochondria, acts downstream of PRKN and PINK1 to promote mitophagy or prevent apoptosis; polyubiquitination by PRKN promotes mitophagy, while monoubiquitination by PRKN decreases mitochondrial calcium influx which ultimately inhibits apoptosis (PubMed:32047033). May participate in the formation of the permeability transition pore complex (PTPC) responsible for the release of mitochondrial products that triggers apoptosis (PubMed:15033708, PubMed:25296756). May mediate ATP export from cells (PubMed:30061676). Part of a complex composed of HSPA9, ITPR1 and VDAC1 that regulates mitochondrial calcium-dependent apoptosis by facilitating calcium transport from the ER lumen to the mitochondria intermembrane space thus providing calcium for the downstream calcium channel MCU that directly releases it into mitochondria matrix (By similarity). Mediates cytochrome c efflux (PubMed:20230784). Catalyzes the scrambling of phospholipids across the outer mitochondrial membrane; the mechanism is unrelated to channel activity and is capable of translocating both anionic and zwitterionic phospholipids.
VDAC3
VDAC, VDAC1, Non-selective voltage-gated ion channel VDAC1, Outer mitochondrial membrane protein porin 1, Plasmalemmal porin, Porin 31HL, Porin 31HM, Voltage-dependent anion-selective channel protein 1, VDAC-1, hVDAC1
Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) is a mouse monoclonal antibody detecting VDAC1/Porin in Western Blot, Flow Cytometry, ICC/IF. Suitable for Cow, Human, Mouse, Rat.
- KO validated for confirmed specificity
- Over 580 publications
- Trusted since 2005
pH: 7.5
Preservative: 0.02% Sodium azide
Constituents: 0.88% Sodium chloride, 0.36% HEPES
Near homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
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.
Product Specifications
Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) is a mouse monoclonal antibody and is validated for use in Flow Cyt, ICC/IF, WB in cow, human, mouse, rat samples.
Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) specifically detects VDAC1/Porin + VDAC3 (UniProt ID: P21796; Molecular weight: 31kDa) and is sold in 100 µg selling sizes.
Quality and Validation
Abcam's high quality validation processes ensure Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) has high sensitivity and specificity.
The specificity of Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) has been confirmed by testing in knockout samples.
Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) has been cited over 584 times in peer reviewed journals and is trusted by the scientific community.
Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) has 51 independent reviews from customers.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
VDAC1/Porin + VDAC3 Western blot staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
ab14734 was shown to react with VDAC1 / Porin in HEK-293T wildtype. Loss of signal was observed when knockout sample Human VDAC1 (Porin) knockout HEK-293T cell lysate ab263839 was used. Wild-type and VDAC1 / Porin knockout samples were subjected to SDS-PAGE. ab14734 and Anti-GAPDH antibody EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] - Mitochondrial Loading Control (ab14734) at 1/1000 dilution
Lane 1: Wild-type Hap1 cell lysate at 20 µg
Lane 2: VDAC1 knockout Hap1 cell lysate at 20 µg
Lane 2: Western blot - Human VDAC1 (Porin) knockout HEK-293T cell line (Human VDAC1 (Porin) knockout HEK-293T cell line ab255444)
Lane 3: Wild-type HEK-293T cell lysate at 20 µg
Lane 4: VDAC1 knockout HEK-293T cell lysate at 20 µg
All lanes: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 17 kDa, 37 kDa
VDAC1/Porin + VDAC3 Western blot staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
Immunoblot analysis of endogenous VDAC1/Porin and VDAC3 in subcellular fractions of MA-10 cell lysates. VDAC1/Porin and VDAC3 are used as a mitochondrial marker proteins.
Lane 1: Whole cell lysate
Lane 2: Cytosolic fraction
Lane 3: Mitochondrial-enriched fraction
All lanes: Western blot - Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] - Mitochondrial Loading Control (ab14734)
VDAC1/Porin + VDAC3 Flow Cytometry staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
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 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (Mouse IgG2b [PLPV219] - Isotype Control 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.
VDAC1/Porin + VDAC3 Immunocytochemistry/ Immunofluorescence staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
ab14734 staining VDAC1/Porin + VDAC3 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.
VDAC1/Porin + VDAC3 Western blot staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
N-GST tagged Recombinant Human VDAC1 protein is available as Recombinant Human VDAC1/Porin protein ab132481
N-GST tagged Recombinant Human VDAC2 protein is available as Recombinant Human VDAC2 protein ab152793
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] - Mitochondrial Loading Control (ab14734) at 1/1000 dilution
Lane 1: N-GST tagged Recombinant Human VDAC1 (aa1 to 283) protein at 0.01 µg
Lane 2: N-GST tagged Recombinant Human VDAC2 (aa 1 to 294) protein at 0.01 µg
Lane 3: N-GST tagged Recombinant Human VDAC3 (aa 1 to 283) protein at 0.01 µg
Observed band size: 33 kDa
Exposure time: 10s
VDAC1/Porin + VDAC3 Western blot staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
Blocking and dilution buffer: 5% NFDM /TBST.
All lanes: Western blot - Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] - Mitochondrial Loading Control (ab14734) at 1/1000 dilution
Lane 1: Wild type HAP1 whole cell lysate at 20 µg
Lane 2: VDAC1 knockout HAP1 whole cell lysate at 20 µg
Observed band size: 33 kDa
Exposure time: 5s
VDAC1/Porin + VDAC3 Western blot staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] - Mitochondrial Loading Control (ab14734) at 1/1000 dilution
Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 2: HAP1 (Wildtype control Human chronic myelogenous leukemia near-haploid cell line) whole cell lysate at 20 µg
Lane 3: MEF (Mus musculus Embryo Fibroblast) whole cell lysate at 20 µg
Lane 4: Rat brain tissue lysate at 20 µg
Observed band size: 33 kDa
Exposure time: 5s
VDAC1/Porin + VDAC3 Western blot staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
Lysates from wild-type MEFs and VDAC1/3-/- MEFs were analyzed by western blotting with Anti-VDAC1/Porin + VDAC3 (top panel) or anti Tom20 antibodies (bottom panel).
(B) The Anti-VDAC1/Porin + VDAC3 antibody does not recognize VDAC2 in VDAC1/3-/- MEFs.
HA-tagged VDAC1, 2 or 3 were expressed in VDAC1/3-/- MEFs, immunoprecipitated with anti-HA antibodies and analyzed by western blotting with anti-HA antibodies (top panel) or Anti-VDAC1/Porin + VDAC3 antibodies (bottom panel). The Anti-VDAC1/Porin + VDAC3 antibodies recognize HA-VDAC1 and HA-VDAC3, but not HA-VDAC2 . Note that HA-VDAC3 migrates at a lower molecular weight than HA-VDAC1 and HA-VDAC2 .
Molecular weights are indicated in kDa.
All lanes: Western blot - Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] - Mitochondrial Loading Control (ab14734)
VDAC1/Porin + VDAC3 Western blot staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
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.
All lanes: Western blot - Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] - Mitochondrial Loading Control (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
VDAC1/Porin + VDAC3 Immunocytochemistry/ Immunofluorescence staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
Immunofluoresence using ab14734 at 0.2 μg/ml on human fibroblasts (red).
Nuclei were labeled with DAPI (blue).
VDAC1/Porin + VDAC3 Western blot staining of MCF7 (Human breast adenocarcinoma cell line) cell lysate using mouse Anti-VDAC1/Porin + VDAC3 antibody
0.5% TBS-tween + Lait 5% NaN3 for 16 hours at 4°C.
All lanes: Western blot - Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] - Mitochondrial Loading Control (ab14734) at 1/500 dilution
All lanes: MCF7 (Human breast adenocarcinoma cell line) cell lysate at 10 µg
All lanes: HRP-conjugated goat anti-mouse polyclonal at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Exposure time: 3min
VDAC1/Porin + VDAC3 Western blot staining using mouse Anti-VDAC1/Porin + VDAC3 antibody
All lanes: Western blot - Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] - Mitochondrial Loading Control (ab14734) at 1/5000 dilution
Lanes 1 - 2: Rat brain cell lysate (homogenate) at 30 µg
Lanes 3 - 4: Rat brain cell lysate (mitochondrial) at 30 µg
All lanes: HRP conjugated sheep anti-mouse IgG
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 39 kDa
Exposure time: 5min
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