Overview

  • Product name
  • Description
    Mouse monoclonal to ATP6V0D1
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant fragment: AKLFPHCGRL YPEGLAQLAR ADDYEQVKNV ADYYPEYKLL FEGAGSNPGD KTLEDRFFEH EVKLNKLAFL N, corresponding to amino acids 238-309 of Human ATP6V0D1

  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab202899 as a replacement.

Properties

Applications

Our Abpromise guarantee covers the use of ab56441 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 - 5 µg/ml. Predicted molecular weight: 40 kDa.
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 10 µg/ml.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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

  • ATP6V0D1 antibody (ab56441) used in immunohistochemistry at 5ug/ml on formalin fixed and paraffin embedded human stomach.


  • Predicted band size : 40 kDa
    ATP6V0D1 antibody (ab56441) at 1ug/lane + HeLa cell lysate at 25ug/lane.
  • ICC/IF image of ab56441 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab56441, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HeLa cells stained with ab56441 (red line). The cells were fixed with 80% methanol (5 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 (ab56441, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was a mix of mouse IgG1 [ICIGG1], (ab91353, 2μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4%PFA/permeabilized in 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:
  • Jacquin E  et al. Pharmacological modulators of autophagy activate a parallel noncanonical pathway driving unconventional LC3 lipidation. Autophagy 13:854-867 (2017). Read more (PubMed: 28296541) »
  • Florey O  et al. V-ATPase and osmotic imbalances activate endolysosomal LC3 lipidation. Autophagy 11:88-99 (2015). Read more (PubMed: 25484071) »

See all 5 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Pig Cell lysate - other (LLC-PK1)
Gel Running Conditions
Non-reduced Denaturing (4-15%)
Loading amount
10 µg
Specification
LLC-PK1
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 1.5% · Temperature: 21°C
Username

Dr. Christina Mcguire

Verified customer

Submitted Dec 22 2015

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxxx.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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