Antibodies, Proteins, Kits and Reagents for Life Science

294 835 ₸ Product size

100 µl 294 835 ₸

Order now and get it on Tuesday March 02, 2021

Anti-ATP6V1A antibody [EPR19270] (ab199326)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19270] to ATP6V1A
Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cyt
Reacts with: Mouse, Rat, Human

Product name

Anti-ATP6V1A antibody [EPR19270]
See all ATP6V1A primary antibodies
Description

Rabbit monoclonal [EPR19270] to ATP6V1A
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Mouse Human
IHC-P	Mouse Rat Human
IP	Mouse 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 heart, fetal liver and fetal kidney lysates; HeLa, K562, HEK-293, C6, PC-12 and NIH/3T3 whole cell lysates; Mouse brain and kidney lysates; Rat brain and kidney lysates. IHC-P: Human kidney, Human thyroid cancer, mouse kidney and rat stomach tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: HeLa cells. IP: HeLa and NIH/3T3 whole cell lysates.
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, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19270
Isotype

IgG
Research areas

Western blot - Anti-ATP6V1A antibody [EPR19270] (ab199326)

All lanes : Anti-ATP6V1A antibody [EPR19270] (ab199326) at 1/2000 dilution

Lane 1 : Human fetal heart lysate
Lane 2 : Human fetal liver lysate
Lane 3 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 68 kDa
Observed band size: 68 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 3 minutes; Lane 2 and 3: 8 seconds.
Western blot - Anti-ATP6V1A antibody [EPR19270] (ab199326)

All lanes : Anti-ATP6V1A antibody [EPR19270] (ab199326) at 1/2000 dilution

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 68 kDa
Observed band size: 68 kDa

Exposure time: 8 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-ATP6V1A antibody [EPR19270] (ab199326)

All lanes : Anti-ATP6V1A antibody [EPR19270] (ab199326) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat kidney lysate
Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 7 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 68 kDa
Observed band size: 68 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 4 seconds; Lane 3 and 4: 1 second; Lane 5,6 and 7: 4 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATP6V1A antibody [EPR19270] (ab199326)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling ATP6V1A with ab199326 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on kidney tubules of the normal Human kidney 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-ATP6V1A antibody [EPR19270] (ab199326)

Immunohistochemical analysis of paraffin-embedded Human thyroid cancer tissue labeling ATP6V1A with ab199326 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on tumor cells of the Human thyroid cancer 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-ATP6V1A antibody [EPR19270] (ab199326)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling ATP6V1A with ab199326 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on kidney tubules of the mouse kidney 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-ATP6V1A antibody [EPR19270] (ab199326)

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling ATP6V1A with ab199326 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasm staining on rat stomach 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-ATP6V1A antibody [EPR19270] (ab199326)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATP6V1A with ab199326 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 Anti-alpha Tubulin antibody [EPR19270]- Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab199326 at 1/250 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-ATP6V1A antibody [EPR19270] (ab199326)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling ATP6V1A with ab199326 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 NIH/3T3 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [EPR19270] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab199326 at 1/250 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Flow Cytometry - Anti-ATP6V1A antibody [EPR19270] (ab199326)

Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATP6V1A with ab199326 at 1/120 dilution (red) compared with a Rabbit IgG,monoclonal -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-ATP6V1A antibody [EPR19270] (ab199326)

ATP6V1A was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab199326 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab199326 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: HeLa whole cell lysate 10µg (Input).

Lane 2: ab199326 IP in HeLa whole cell lysate.

Lane 3: Rabbit IgG,monoclonal [EPR19270] - Isotype Control (ab172730) instead of ab199326 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 5 seconds.
Immunoprecipitation - Anti-ATP6V1A antibody [EPR19270] (ab199326)

ATP6V1A was immunoprecipitated from 1mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab199326 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab199326 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: NIH/3T3 whole cell lysate 10µg (Input).

Lane 2: ab199326 IP in NIH/3T3 whole cell lysate.

Lane 3: Rabbit IgG,monoclonal [EPR19270] - Isotype Control (ab172730) instead of ab199326 in NIH/3T3 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 5 seconds.
Anti-ATP6V1A antibody [EPR19270] (ab199326)

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