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Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR10852(B)] to VDAC1 / Porin - Mitochondrial Loading Control
Suitable for: WB, IHC-P, ICC/IF, IHC-Fr
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control
See all VDAC1 / Porin primary antibodies
Description

Rabbit monoclonal [EPR10852(B)] to VDAC1 / Porin - Mitochondrial Loading Control
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-Fr	Mouse
IHC-P	Mouse Rat Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide within Human VDAC1/ Porin aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P21796
Positive control
- WB: HepG2, Jurkat, HEK-293, HAP1 and HeLa cell lysates; Mouse and rat kidney lysate; Rat cerebellum whole tissue lysate IHC-P: Human liver, heart, kidney, ovarian carcinoma, thyroid gland carcinoma, skeletal muscle and cervical carcinoma tissues; Rat kidney tissue; Mouse cardiac muscle tissue; ICC/IF: HeLa and Jurkat cells; IHC-Fr: Mouse cardiac and skeletal muscle tissues.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

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, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR10852(B)
Isotype

IgG
Research areas

Western blot - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

All lanes : Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : VDAC1 knockout HAP1 whole cell lysate
Lane 3 : HEK-293T whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 31 kDa
Observed band size: 31 kDa

Lanes 1 - 3: Merged signal (red and green). Green - ab154856 observed at 31 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab154856 was shown to specifically react with in wild-type HAP1 cells as signal was lost in VDAC1 knockout cells. Wild-type and VDAC1 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab154856 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Frozen sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemistry (Frozen sections) analysis of mouse skeletal muscle tissue sections labeling VDAC1 / Porin with Purified ab154856 at 1/50 (0.7 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab154856 at a working dilution of 1/200. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

All lanes : Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856) 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 HEK-293T knockout cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 31 kDa
Observed band size: 32 kDa
why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab154856 observed at 31 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab154856 was shown to react with VDAC1 / Porin in wild-type HEK-293T. 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. ab154856 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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 (ab216773) and 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.
Western blot - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

All lanes : Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856) at 1/10000 dilution (purified)

Lane 1 : HepG2 cell lysate
Lane 2 : HEK293 cell lysate
Lane 3 : HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 31 kDa
Observed band size: 31 kDa

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunocytochemistry/ Immunofluorescence - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

ab154856 staining VDAC1 / Porin showing cytoplasmic staining in HeLa cells (Human cervix adenocarcinoma epithelial cells) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol, Samples were incubated with primary antibody (1/1000) for 1 hour at 21°C. ab150077, an Alexa Fluor® 488-conjugated Goat anti-Rabbit IgG (1:1000) was used as the secondary antibody. DAPI (1/200) was used as a counter stain.
Western blot - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

All lanes : Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856) at 1/10000 dilution (purified)

Lane 1 : mouse kidney lysate
Lane 2 : rat kidney lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 31 kDa
Observed band size: 31 kDa

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Frozen sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemistry (Frozen sections) analysis of mouse cardiac muscle tissue sections labeling VDAC1 / Porin with Purified ab154856 at 1/50 (0.7 µg/ml).Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. DAPI was used as a counterstain.
Western blot - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

All lanes : Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856) at 1/1000 dilution (unpurified)

Lane 1 : HepG2 cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : 293T cell lysate
Lane 4 : HeLa cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 31 kDa

Secondary antibody - anti-rabbit HRP (ab6721)
Immunocytochemistry/ Immunofluorescence - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

ab154856 staining VDAC1 / Porin showing cytoplasmic staining in Jurkat cells (Human T cell leukemia T lymphocyte) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol, Samples were incubated with primary antibody (1/1000) for 1 hour at 21°C. ab150077, an Alexa Fluor® 488-conjugated Goat anti-Rabbit IgG (1:1000) was used as the secondary antibody. DAPI (1/200) was used as a counter stain.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemical staining of paraffin embedded rat kidney with purified ab154856 at a working dilution of 1/200. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856) at 1/2000 dilution (purified) + Jurkat cell lysate at 20 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 31 kDa
Observed band size: 31 kDa

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemical staining of paraffin embedded mouse cardiac muscle with purified ab154856 at a working dilution of 1/200. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Western blot - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856) This image is courtesy of an anonymous Abreview

Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856) at 1/5000 dilution (unpurified) + Rat cerebellum whole tissue lysate at 30 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97069) (undiluted)

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 31 kDa
Observed band size: 31 kDa

Exposure time: 2 seconds
See Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling VDAC1 with unpurified ab154856 at 1/100 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemical analysis of paraffin-embedded human heart tissue labeling VDAC1 with unpurified ab154856 at 1/100 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemical analysis of paraffin embedded human normal kidney tissue using unpurified ab154856 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemical analysis of paraffin embedded human ovarian carcinoma tissue using unpurified ab154856 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemical analysis of paraffin embedded human thyroid gland carcinoma tissue using unpurified ab154856 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Immunohistochemical analysis of paraffin embedded human skeletal muscle tissue using unpurified ab154856 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

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Anti-VDAC1 / Porin antibody [EPR10852(B)] - Mitochondrial Loading Control (ab154856)

Key features and details

Overview

Product name

Description

Host species

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Images

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