Overview

  • Product name

    Anti-ATP6V1A antibody [EPR19271]
    See all ATP6V1A primary antibodies
  • Description

    Rabbit monoclonal [EPR19271] to ATP6V1A
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human ATP6V1A aa 450 to the C-terminus. The exact sequence is proprietary.
    Database link: P38606

  • Positive control

    • WB: Human fetal liver, fetal heart and fetal kidney lysates; HeLa, K562, HEK-293, C6, PC-12, NIH/3T3 and RAW 264.7 whole cell lysates; Mouse brain, kidney and spleen lysates; Rat brain, kidney and spleen lysates. IHC-P: Human kidney and breast cancer tissues; Mouse cerebrum and rat kidney tissues. IP: HeLa whole cell lysate.
  • General notes

     

     

    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

Applications

Our Abpromise guarantee covers the use of ab199325 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
IP 1/50.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).

Target

  • Function

    Catalytic subunit of the peripheral V1 complex of vacuolar ATPase. V-ATPase vacuolar ATPase is responsible for acidifying a variety of intracellular compartments in eukaryotic cells.
  • Tissue specificity

    Present in all tissues analyzed.
  • Sequence similarities

    Belongs to the ATPase alpha/beta chains family.
  • Information by UniProt
  • Database links

  • Alternative names

    • 70kDa subunit antibody
    • ATP6A1 antibody
    • ATP6V1A antibody
    • ATP6V1A1 antibody
    • ATPase H+ transporting lysosomal subunit A1 antibody
    • ATPase H+ transporting, lysosomal 70kDa V1 subunit A antibody
    • H(+) transporting two sector ATPase subunit A antibody
    • H+ transporting ATPase chain A vacuolar (VA68 type) antibody
    • HO68 antibody
    • OTTHUMP00000214746 antibody
    • V ATPase 69 kDa subunit 1 antibody
    • V ATPase A subunit 1 antibody
    • V ATPase subunit A 1 antibody
    • V-ATPase 69 kDa subunit antibody
    • V-ATPase subunit A antibody
    • V-type proton ATPase catalytic subunit A antibody
    • VA68 antibody
    • Vacuolar ATP synthase catalytic subunit A ubiquitous isoform antibody
    • Vacuolar ATPase isoform VA68 antibody
    • Vacuolar proton pump alpha subunit 1 antibody
    • Vacuolar proton pump subunit alpha antibody
    • VATA_HUMAN antibody
    • Vma1 antibody
    • VPP2 antibody
    see all

Images

  • All lanes : Anti-ATP6V1A antibody [EPR19271] (ab199325) at 1/2000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 68 kDa
    Observed band size: 68 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 2: 8 seconds; Lane 3: 3 seconds

  • All lanes : Anti-ATP6V1A antibody [EPR19271] (ab199325) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 68 kDa
    Observed band size: 68 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-ATP6V1A antibody [EPR19271] (ab199325) at 1/2000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat brain lysate
    Lane 5 : Rat kidney lysate
    Lane 6 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 68 kDa
    Observed band size: 68 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1 and 2: 1 second; Lane 3: 3 seconds; Lane 4 and 5: 1 second; Lane 6: 3 seconds.

     

  • All lanes : Anti-ATP6V1A antibody [EPR19271] (ab199325) at 1/2000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 68 kDa
    Observed band size: 68 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1,2 and 3: 3 seconds; Lane 4: 1 second.

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling ATP6V1A with ab199325 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on kidney tubules of normal Human kidney 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 cancer tissue labeling ATP6V1A with ab199325 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on cancer cells of the breast 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 cerebrum tissue labeling ATP6V1A with ab199325 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on neurons of the mouse cerebrum 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 ATP6V1A with ab199325 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasm staining on kidney tubules of the rat kidney 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.

  • ATP6V1A was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab199325 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab199325 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
    Lane 1: HeLa whole cell lysate 10µg (Input).
    Lane 2: ab199325 IP in HeLa whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199325 in HeLa whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 seconds.

References

ab199325 has not yet been referenced specifically in any publications.

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