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

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: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-Pmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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 (intra): HeLa cells. IP: HeLa whole cell lysate.
General notes
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.

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
Isotype

IgG
Research areas

Alternative Versions
- Anti-ATP6V0D1/P39 antibody [EPR18320] - BSA and Azide free (ab251385)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
Recombinant Protein
- Recombinant Human ATP6V0D1/P39 protein (ab160342)

The Abpromise guarantee

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
Flow Cyt (Intra)		1/100.
WB		1/2000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
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.

Notes
Flow Cyt (Intra) 1/100.
WB 1/2000. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
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.

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.
Target information above from: UniProt accession P61421 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
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

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

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.
Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

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

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

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
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.
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.
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.
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.
Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

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.
Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

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

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) labeling ATP6V0D1/P39with 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.
Immunoprecipitation - Anti-ATP6V0D1/P39 antibody [EPR18320] (ab202897)

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

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

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Publishing research using ab202897? Please let us know so that we can cite the reference in this datasheet.

ab202897 has been referenced in 1 publication.

Kanfer G et al. Image-based pooled whole-genome CRISPRi screening for subcellular phenotypes. J Cell Biol 220:N/A (2021). PubMed: 33464298

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (1)

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