Anti-VDAC1/Porin + VDAC3 抗体 [20B12AF2] (ab14734)
Key features and details
- Mouse monoclonal [20B12AF2] to VDAC1/Porin + VDAC3
- Suitable for: WB, ICC/IF, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Cow, Human
- Isotype: IgG2b
製品の概要
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製品名
Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] -
製品の詳細
Mouse monoclonal [20B12AF2] to VDAC1/Porin + VDAC3 -
由来種
Mouse -
アプリケーション
適用あり: WB, ICC/IF, Flow Cytmore details -
種交差性
交差種: Mouse, Rat, Cow, Human
交差が予測される動物種: Sheep, Goat, Cat, Dog, Pig, Drosophila melanogaster, Fish, Quail, Common marmoset, Dogfish, Catshark -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: Isolated mitochondria from human, cow, rat and mouse heart. HepG2 cell lysate. ICC/IF: HeLa cells. Human fibroblasts. Flow Cyt: HepG2 cells.
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特記事項
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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
バッファー
pH: 7.50
Preservative: 0.02% Sodium azide
Constituents: 0.36% HEPES, 0.88% Sodium chloride -
Concentration information loading...
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精製度
IgG fraction -
特記事項(精製)
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. -
ポリ/モノ
モノクローナル -
クローン名
20B12AF2 -
アイソタイプ
IgG2b -
軽鎖の種類
kappa -
研究分野
関連製品
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab14734の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB | (18) |
Use a concentration of 1 µg/ml. Detects a band of approximately 39 kDa.
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ICC/IF | (4) |
Use at an assay dependent concentration.
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Flow Cyt |
Use 1µg for 106 cells.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
特記事項 |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 39 kDa. |
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. |
ターゲット情報
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細胞内局在
VDAC1/Porin: Mitochondrion outer membrane. Cell membrane. VDAC3: Mitochondrial outer membrane. -
参照データベース
- Entrez Gene: 282119 Cow
- Entrez Gene: 282716 Cow
- Entrez Gene: 7416 Human
- Entrez Gene: 7419 Human
- Entrez Gene: 22333 Mouse
- Entrez Gene: 22335 Mouse
- Entrez Gene: 397010 Pig
- Entrez Gene: 397651 Pig
see all
画像
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All lanes : Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) at 1/1000 dilution
Lane 1 : Wild-type Hap1 cell lysate
Lane 2 : VDAC1 knockout Hap1 cell lysate
Lane 3 : Wild-type HEK-293T cell lysate
Lane 4 : VDAC1 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.Lanes 1 - 4: Merged signal (red and green). Green - ab14734 observed at 31 kDa. Red - loading control, ab181602 observed at 37 kDa. The lower band very close to VDAC1 is likely to be VDAC3.
ab14734 was shown to react with VDAC1/Porin + VDAC3 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255444 (knockout 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 (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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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.
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All lanes : Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) at 1/1000 dilution
Lane 1 : N-GST tagged Recombinant Human VDAC1 (aa1 to 283) protein with NFDM/TBST
Lane 2 : N-GST tagged Recombinant Human VDAC2 (aa 1 to 294) protein with NFDM/TBST
Lane 3 : N-GST tagged Recombinant Human VDAC3 (aa 1 to 283) protein with NFDM/TBST
Lysates/proteins at 0.01 µg per lane.
Blocking peptides at 5 % per lane.
Observed band size: 33 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsDiluting buffer: 5% NFDM /TBST
N-GST tagged Recombinant Human VDAC1 protein is available as ab132481
N-GST tagged Recombinant Human VDAC2 protein is available as ab152793
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All lanes : Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) at 1/1000 dilution
Lane 1 : Wild type HAP1 whole cell lysate
Lane 2 : VDAC1 knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Observed band size: 33 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking and dilution buffer: 5% NFDM /TBST.
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All lanes : Anti-VDAC1/Porin + VDAC3 antibody [20B12AF2] (ab14734) at 1/1000 dilution
Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate with NFDM/TBST
Lane 2 : HAP1 (Wildtype control Human chronic myelogenous leukemia near-haploid cell line) whole cell lysate with NFDM/TBST
Lane 3 : MEF (Mus musculus Embryo Fibroblast) whole cell lysate with NFDM/TBST
Lane 4 : Rat brain tissue lysate with NFDM/TBST
Lysates/proteins at 20 µg per lane.
Blocking peptides at 5 % per lane.
Observed band size: 33 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsDiluting buffer: 5% NFDM /TBST
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(A) The Anti-VDAC1/Porin + VDAC3 antibody recognizes VDAC1 and VDAC3 but not VDAC2.
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.
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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
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All lanes : Anti-VDAC1/Porin + VDAC3 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. -
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.
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Immunofluoresence using ab14734 at 0.2 µg/ml on human fibroblasts (red).
Nuclei were labeled with DAPI (blue). -
Anti-VDAC1/Porin + VDAC3 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 minutes0.5% TBS-tween + Lait 5% NaN3 for 16 hours at 4ºC.
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All lanes : Anti-VDAC1/Porin + VDAC3 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
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (584)
ab14734 は 584 報の論文で使用されています。
- Kosik B et al. Actovegin improves skeletal muscle mitochondrial respiration and functional aerobic capacity in a type 1 diabetic male murine model. Appl Physiol Nutr Metab 49:265-272 (2024). WB ; Mouse . PubMed: 37913525
- Janer A et al. ESYT1 tethers the ER to mitochondria and is required for mitochondrial lipid and calcium homeostasis. Life Sci Alliance 7:N/A (2024). PubMed: 37931956
- Potes Y et al. Melatonin Alleviates the Impairment of Muscle Bioenergetics and Protein Quality Control Systems in Leptin-Deficiency-Induced Obesity. Antioxidants (Basel) 12:N/A (2023). PubMed: 38001815
- Yang Y et al. Cyclophilin D-induced mitochondrial impairment confers axonal injury after intracerebral hemorrhage in mice. Neural Regen Res 18:849-855 (2023). PubMed: 36204853
- Gingerich MA et al. An intrinsically disordered protein region encoded by the human disease gene CLEC16A regulates mitophagy. Autophagy 19:525-543 (2023). PubMed: 35604110