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

This data was developed using ab199326, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times: Lane 1: 3 minutes; Lanes 2-3: 8 seconds.

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

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

This data was developed using ab199326, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

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 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

This data was developed using ab199326, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times: Lanes 1-2: 4 seconds; Lanes 3-4: 1 second; Lanes 5-7: 4 seconds.

This data was developed using ab199326, the same antibody clone in a different buffer formulation.

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.

This data was developed using ab199326, the same antibody clone in a different buffer formulation.

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.

This data was developed using ab199326, the same antibody clone in a different buffer formulation.

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.

This data was developed using ab199326, the same antibody clone in a different buffer formulation.

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.

This data was developed using ab199326, the same antibody clone in a different buffer formulation.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.

This data was developed using ab199326, the same antibody clone in a different buffer formulation.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.

This data was developed using ab199326, the same antibody clone in a different buffer formulation.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.

This data was developed using ab199326, the same antibody clone in a different buffer formulation.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.

This data was developed using ab199326, the same antibody clone in a different buffer formulation.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.