Recombinant
RabMAb

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

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

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

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

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 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

  • 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

  • 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.

  • 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

  • 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

  • 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

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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 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.

  • 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.

References

ab202897 has not yet been referenced specifically in any publications.

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