Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18320-38] to ATP6V0D1/P39
  • Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ATP6V0D1/P39 antibody [EPR18320-38]
    See all ATP6V0D1/P39 primary antibodies

  • Description

    Rabbit monoclonal [EPR18320-38] to ATP6V0D1/P39

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Mouse
    Rat
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human fetal kidney, Human fetal brain, HeLa, MCF-7, A431, C6, PC-12 and PC-12 lysates. Mouse kidney and spleen lysates. Rat spleen lysate. IHC-P: Human breast and cervix carcinoma tissue. Mouse and Rat kidney tissue. ICC/IF: HeLa and MCF-7 cells. IP: MCF-7 whole cell lysate. Flow Cyt: HeLa cells.

  • 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.

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

    EPR18320-38

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)
    Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)
    All lanes : Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899) at 1/10000 dilution

    Lane 1 : Human fetal kidney
    Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma)
    Lane 3 : MCF-7 (Human breast adenocarcinoma cell line)
    Lane 4 : A431 (Human epidermoid carcinoma)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (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 [EPR18320-38] (ab202899)
    Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)
    Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899) at 1/2000 dilution + Human fetal brain at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

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


    Exposure time: 5 seconds


    Blocking/dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)
    Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)
    All lanes : Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899) at 1/2000 dilution

    Lane 1 : Mouse kidney
    Lane 2 : Mouse spleen
    Lane 3 : Rat spleen
    Lane 4 : C6 (Rat glial tumor cells)
    Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma)
    Lane 6 : NIH/3T3 (mouse embryo fibroblast cells)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (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 [EPR18320-38] (ab202899)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

    Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 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.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

    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 [EPR18320-38] (ab202899)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

    Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 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.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

    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 [EPR18320-38] (ab202899)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

    Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 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.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

    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 [EPR18320-38] (ab202899)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

    Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 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.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)
    Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ATP6V0D1/P39 with ab202899 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasm staining on HeLa cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab202899 at 1/500 dilution followed by ab150120 (Alexa Fluor® 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 [EPR18320-38] (ab202899)
    Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF-7 (Human breast adenocarcinoma) cells labeling ATP6V0D1/P39 with ab202899 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasm staining on MCF-7 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab202899 at 1/500 dilution followed byab150120 (Alexa Fluor® 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 [EPR18320-38] (ab202899)
    Flow Cytometry - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

    Flow Cytometry analysis of HeLa cells labelling ATP6V0D1/P39 with ab202899 at 1/800 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunoprecipitation - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)
    Immunoprecipitation - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

    ATP6V0D1/P39 was immunoprecipitated from 1mg of MCF-7 (Human breast adenocarcinoma cell line) whole cell lysate with ab202899 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab202899 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: Input MCF-7 whole cell lysate (10 µg). Lane 2: MCF-7 whole cell lysate following precipitation. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202899 in MCF-7 whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.10 second exposure.

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