RecombinantRabMAb

Recombinant Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18320] to ATP6V0D1/P39
  • Suitable for: WB, Flow Cyt, ICC/IF, IP, IHC-P
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ATP6V0D1/P39 antibody [EPR18320]
    See all ATP6V0D1/P39 primary antibodies

  • Description

    Rabbit monoclonal [EPR18320] to ATP6V0D1/P39

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IF, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human ATP6V0D1/P39 aa 100-350. The exact sequence is proprietary.
    Database link: P61421

  • Positive control

    • WB: Human fetal kidney, fetal brain, fetal heart and breast cancer lysates; HeLa, MCF7, A431, C6, RAW 264.7, PC-12 and NIH/3T3 cell lysates; Mouse brain, mouse kidney and rat kidney lysates. IHC-P: Human cervix carcinoma, Human breast carcinoma, mouse kidney and rat kidney tissue. ICC/IF: HeLa and MCF7 cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.

  • General notes

     

     

     This product was previously labelled as ATP6V0D1

     

    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.

    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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR18320

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab202897 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 1/2000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
Flow Cyt 1/100.
ICC/IF 1/250.
IP 1/40.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Subunit of the integral membrane V0 complex of vacuolar ATPase. Vacuolar ATPase is responsible for acidifying a variety of intracellular compartments in eukaryotic cells, thus providing most of the energy required for transport processes in the vacuolar system. May play a role in coupling of proton transport and ATP hydrolysis (By similarity). May play a role in cilium biogenesis through regulation of the transport and the localization of proteins to the cilium.

  • Tissue specificity

    Ubiquitous.

  • Sequence similarities

    Belongs to the V-ATPase V0D/AC39 subunit family.

  • Cellular localization

    Membrane. Localizes to centrosome and the base of the cilium.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 32 kDa accessory protein antibody
    • ATP6D antibody
    • ATP6DV antibody
    • ATP6V0D1 antibody
    • ATPase H+ transporting lysosomal (vacuolar proton pump) member D antibody
    • ATPase H+ transporting lysosomal 38kD V0 subunit d antibody
    • ATPase H+ transporting lysosomal 38kDa V0 subunit d1 antibody
    • ATPase H+ transporting lysosomal V0 subunit d1 antibody
    • H(+) transporting two sector ATPase subunit D antibody
    • p39 antibody
    • V ATPase 40 KDa accessory protein antibody
    • V ATPase AC39 subunit antibody
    • V ATPase subunit d 1 antibody
    • V ATPase subunit D antibody
    • V-ATPase 40 kDa accessory protein antibody
    • V-ATPase AC39 subunit antibody
    • V-ATPase subunit d 1 antibody
    • V-type proton ATPase subunit d 1 antibody
    • VA0D1_HUMAN antibody
    • Vacuolar ATP synthase subunit d 1 antibody
    • Vacuolar proton pump subunit d 1 antibody
    • VATX antibody
    • VMA 6 antibody
    • VMA6 antibody
    • VPATPD antibody
    see all

Images

  • Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    All lanes : Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897) at 1/10000 dilution

    Lane 1 : Human fetal kidney lysate
    Lane 2 : HeLa (Human eithelial cells from cervix adenocarcinoma) cell lysate
    Lane 3 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
    Lane 4 : A431 (Human epidermoid carcinoma) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 40 kDa
    Observed band size: 40 kDa


    Exposure time: 1 minute


    Blocking/dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    All lanes : Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897) at 1/2000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

    Predicted band size: 40 kDa
    Observed band size: 40 kDa


    Exposure time: 10 seconds


    Blocking/dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897) at 1/2000 dilution + Human breast cancer lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

    Predicted band size: 40 kDa
    Observed band size: 40 kDa


    Exposure time: 3 minutes


    Blocking/dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    All lanes : Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Rat kidney lysate
    Lane 4 : C6 (Rat glial tumor cells) cell lysate
    Lane 5 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cell lysate
    Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma) cell lysate
    Lane 7 : NIH/3T3 (Mouse embryo fibroblast cells) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 40 kDa
    Observed band size: 40 kDa


    Exposure time: 5 seconds


    Blocking/dilution buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

    Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling ATP6V0D1/P39 with ab202897 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling ATP6V0D1/P39 with ab202897 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Human breast carcinoma tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

    Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling ATP6V0D1/P39 with ab202897 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse kidney tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

    Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling ATP6V0D1/P39 with ab202897 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on Rat kidney tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ATP6V0D1/P39 with ab202897 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:-
    -ve control 1: - ab202897 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling ATP6V0D1/P39 with ab202897 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on MCF7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:-
    -ve control 1: - ab202897 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Flow Cytometry - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Flow Cytometry - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) labeling ATP6V0D1/P39 with ab202897 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Immunoprecipitation - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

    ATP6V0D1/P39 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab202897 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab202897 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: HeLa whole cell lysate 10ug (Input). Lane 2: ab202897 IP in HeLa whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202897 in HeLa whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

  • Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
    Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

References (0)

Publishing research using ab202897? Please let us know so that we can cite the reference in this datasheet.

ab202897 has not yet been referenced specifically in any publications.

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