
Recombinant Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)
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- References
- Protocols
Overview
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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.
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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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18320 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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 |
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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
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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
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Database links
- Entrez Gene: 9114 Human
- Entrez Gene: 11972 Mouse
- Entrez Gene: 291969 Rat
- Omim: 607028 Human
- SwissProt: P61421 Human
- SwissProt: P51863 Mouse
- Unigene: 106876 Human
- Unigene: 17708 Mouse
see all -
Alternative names
- 32 kDa accessory protein antibody
- ATP6D antibody
- ATP6DV antibody
see all
Images
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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 minuteBlocking/dilution buffer: 5% NFDM/TBST.
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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 secondsBlocking/dilution buffer: 5% NFDM/TBST
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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 minutesBlocking/dilution buffer: 5% NFDM/TBST
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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 secondsBlocking/dilution buffer: 5% NFDM/TBST
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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.
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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.
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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.
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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.
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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.
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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.
Protocols
Datasheets and documents
References
ab202897 has not yet been referenced specifically in any publications.