Antibodies, Proteins, Kits and Reagents for Life Science

284 784 ₸ Product size

100 µl 284 784 ₸

Order now and get it on Tuesday March 02, 2021

Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18320-38] to ATP6V0D1/P39
Suitable for: Flow Cyt, WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-ATP6V0D1/P39 antibody [EPR18320-38]
See all ATP6V0D1/P39 primary antibodies
Description

Rabbit monoclonal [EPR18320-38] 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, Human fetal brain, HeLa, MCF-7, A431, C6, PC-12 and PC-12 lysates. Mouse kidney and spleen lysates. Rat spleen lysate. IHC-P: Human breast and cervix carcinoma tissue. Mouse and Rat kidney tissue. ICC/IF: HeLa and MCF-7 cells. IP: MCF-7 whole cell lysate. Flow Cyt: HeLa cells.
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

EPR18320-38
Isotype

IgG
Research areas

Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

All lanes : Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899) at 1/10000 dilution

Lane 1 : Human fetal kidney
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma)
Lane 3 : MCF-7 (Human breast adenocarcinoma cell line)
Lane 4 : A431 (Human epidermoid carcinoma)

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (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 [EPR18320-38] (ab202899)

Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899) at 1/2000 dilution + Human fetal brain at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 40 kDa
Observed band size: 40 kDa

Exposure time: 5 seconds

Blocking/dilution buffer: 5% NFDM/TBST.
Western blot - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

All lanes : Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899) at 1/2000 dilution

Lane 1 : Mouse kidney
Lane 2 : Mouse spleen
Lane 3 : Rat spleen
Lane 4 : C6 (Rat glial tumor cells)
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma)
Lane 6 : NIH/3T3 (mouse embryo fibroblast cells)

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (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 [EPR18320-38] (ab202899)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 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.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

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 [EPR18320-38] (ab202899)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 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.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

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 [EPR18320-38] (ab202899)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 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.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

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 [EPR18320-38] (ab202899)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling ATP6V0D1/P39 with ab202899 at 1/250 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.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ATP6V0D1/P39 with ab202899 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasm staining on HeLa cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1 - ab202899 at 1/500 dilution followed by ab150120 (Alexa Fluor^® 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 [EPR18320-38] (ab202899)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF-7 (Human breast adenocarcinoma) cells labeling ATP6V0D1/P39 with ab202899 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasm staining on MCF-7 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1 - ab202899 at 1/500 dilution followed byab150120 (Alexa Fluor^® 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 [EPR18320-38] (ab202899)

Flow Cytometry analysis of HeLa cells labelling ATP6V0D1/P39 with ab202899 at 1/800 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunoprecipitation - Anti-ATP6V0D1/P39 antibody [EPR18320-38] (ab202899)

ATP6V0D1/P39 was immunoprecipitated from 1mg of MCF-7 (Human breast adenocarcinoma cell line) whole cell lysate with ab202899 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab202899 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: Input MCF-7 whole cell lysate (10 µg). Lane 2: MCF-7 whole cell lysate following precipitation. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202899 in MCF-7 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.10 second exposure.

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