Antibodies, Proteins, Kits and Reagents for Life Science

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 & 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, 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.
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.

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

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

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.
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)