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    vdac1--porin-antibody-20b12af2-ab14734.pdf

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Validated using a knockout cell line

Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)

  • Datasheet
  • SDS
Reviews (49)Q&A (25)References (330)

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Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
  • Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
  • Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
  • Immunocytochemistry/ Immunofluorescence - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
  • Flow Cytometry - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
  • Immunocytochemistry/ Immunofluorescence - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
  • Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
  • Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)

Key features and details

  • Mouse monoclonal [20B12AF2] to VDAC1 / Porin
  • Suitable for: WB, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Cow, Human
  • Isotype: IgG2b

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Overview

  • Product name

    Anti-VDAC1 / Porin antibody [20B12AF2]
    See all VDAC1 / Porin primary antibodies
  • Description

    Mouse monoclonal [20B12AF2] to VDAC1 / Porin
  • Host species

    Mouse
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Cow, Human
    Predicted to work with: Sheep, Goat, Cat, Dog, Pig, Drosophila melanogaster, Fish, Quail, Common marmoset, Dogfish, Catshark
  • Immunogen

    Recombinant full length protein. This information is considered to be commercially sensitive.

  • Positive control

    • WB: Isolated mitochondria from human, cow, rat and mouse heart. HepG2 cell lysate. ICC/IF: HeLa cells. Human fibroblasts. Flow Cyt: HepG2 cells.
  • General notes

    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.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: HEPES, Sodium chloride
  • Concentration information loading...
  • Purity

    IgG fraction
  • Purification notes

    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.
  • Clonality

    Monoclonal
  • Clone number

    20B12AF2
  • Isotype

    IgG2b
  • Light chain type

    kappa
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • Mitochondria
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Isotype/Loading Controls
    • Loading Controls
    • VDAC1 - Mitochondrial Preps
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • KO cell lines

    • Human VDAC1 (Porin) knockout HEK293T cell line (ab255444)
  • KO cell lysates

    • Human VDAC1 (Porin) knockout HEK293T cell lysate (ab263839)
  • Positive Controls

    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

Applications

Our Abpromise guarantee covers the use of ab14734 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 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.

Target

  • Function

    Forms 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 specificity

    Heart, liver and skeletal muscle.
  • Sequence similarities

    Belongs to the eukaryotic mitochondrial porin family.
  • Domain

    Consists 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 localization

    Mitochondrion outer membrane. Cell membrane.
  • Target information above from: UniProt accession P21796 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 282119 Cow
    • Entrez Gene: 7416 Human
    • Entrez Gene: 22333 Mouse
    • Entrez Gene: 397010 Pig
    • Entrez Gene: 83529 Rat
    • Omim: 604492 Human
    • SwissProt: P45879 Cow
    • SwissProt: P21796 Human
    • SwissProt: Q60932 Mouse
    • SwissProt: Q9MZ16 Pig
    • SwissProt: Q9Z2L0 Rat
    • Unigene: 519320 Human
    • Unigene: 709372 Human
    • Unigene: 3555 Mouse
    • Unigene: 470023 Mouse
    • Unigene: 54594 Rat
    see all
  • Alternative names

    • hVDAC1 antibody
    • mVDAC5 antibody
    • OMP2 antibody
    • Outer mitochondrial membrane protein porin 1 antibody
    • Plasmalemmal porin antibody
    • Porin 31HL antibody
    • Porin 31HM antibody
    • Porin antibody
    • VDAC 1 antibody
    • VDAC 5 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

Images

  • Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
    Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
    All lanes : Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)

    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.  

     ab14734 was shown to react with VDAC1 / Porin 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-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 [20B12AF2] (ab14734)
    Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)Li et al PLoS One. 2016 Feb 22;11(2):e0149728. doi: 10.1371/journal.pone.0149728. eCollection 2016. Fig 5.

    Double immunofluorescent staining and immunoblot analysis of the endogenous SCPX and SCP2 proteins in MA-10 cells.

    Immunoblot analysis of endogenous SCP2 in subcellular fractions of MA-10 cell lysates. VDAC1 is used as a mitochondrial marker protein, and GAPDH is used as a whole cell lysate loading control.

    Lane 1: Whole cell lysate

    Lane 2: Cytosolic fraction

    Lane 3: Mitochondrial-enriched fraction

  • Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
    Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
    All lanes : Anti-VDAC1 / Porin 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.
  • Immunocytochemistry/ Immunofluorescence - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
    Immunocytochemistry/ Immunofluorescence - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)This image is courtesy of an Abreview by Michiel Krols.

    ab14734 staining VDAC1 / Porin 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.

    See Abreview

  • Flow Cytometry - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
    Flow Cytometry - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
    Immunocytochemistry/ Immunofluorescence - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)

    Immunofluoresence using ab14734 at 0.2 µg/ml on human fibroblasts (red).
    Nuclei were labeled with DAPI (blue).

  • Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
    Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)Image courtesy of an anonymous abreview.
    Anti-VDAC1 / Porin 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 minutes


    0.5% TBS-tween + Lait 5% NaN3 for 16 hours at 4ºC.

    See Abreview

  • Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)
    Western blot - Anti-VDAC1 / Porin antibody [20B12AF2] (ab14734)This image is courtesy of an anonymous Abreview
    All lanes : Anti-VDAC1 / Porin 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

    See Abreview

Protocols

  • Mouse on Mouse staining protocol

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (330)

    Publishing research using ab14734? Please let us know so that we can cite the reference in this datasheet.

    ab14734 has been referenced in 330 publications.

    • Dirks ML  et al. Short-term bed rest-induced insulin resistance cannot be explained by increased mitochondrial H2 O2 emission. J Physiol 598:123-137 (2020). WB ; Human . PubMed: 31721213
    • Pignanelli C  et al. Low-load resistance training to task failure with and without blood flow restriction: muscular functional and structural adaptations. Am J Physiol Regul Integr Comp Physiol 318:R284-R295 (2020). WB ; Human . PubMed: 31823670
    • Ke Q  et al. UCP2-induced hypoxia promotes lipid accumulation and tubulointerstitial fibrosis during ischemic kidney injury. Cell Death Dis 11:26 (2020). PubMed: 31932578
    • Lobo-Jarne T  et al. Multiple pathways coordinate assembly of human mitochondrial complex IV and stabilization of respiratory supercomplexes. EMBO J N/A:e103912 (2020). PubMed: 32511785
    • Kathuria A  et al. Transcriptome analysis and functional characterization of cerebral organoids in bipolar disorder. Genome Med 12:34 (2020). PubMed: 32306996
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-10 of 74 Abreviews or Q&A

    Western blot abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

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    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    10 µg
    Specification
    HEK293
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 13 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
    Permeabilization
    No
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 12 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

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    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
    Permeabilization
    No
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More

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    Verified customer

    Submitted Aug 12 2020

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 30 2020

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Pig Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 25 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Zebrafish Tissue sections (Embryo)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Embryo
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 25 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rabbit Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Liver
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 25 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

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    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (B6 Kidney primary culture)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    B6 Kidney primary culture
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 20 2020

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 20 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-VDAC1 / Porin antibody [20B12AF2]

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    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Kidney H9 Organoid)
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney H9 Organoid
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

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    Submitted Feb 06 2020

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