重组Anti-ATP6V1A抗体[EPR19270]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ATP6V1A antibody [EPR19270] (AB199326)

Intracellular 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.

• ICC/IF

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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.

• IHC-P

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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.

• IHC-P

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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.

• IP

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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.

All lanes:

Immunoprecipitation - Anti-ATP6V1A antibody [EPR19270] (ab199326)

Predicted band size: 68 kDa

false

• IHC-P

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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.

• ICC/IF

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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.

• IHC-P

Supplier Data

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.

• IP

Supplier Data

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.

All lanes:

Immunoprecipitation - Anti-ATP6V1A antibody [EPR19270] (ab199326)

Predicted band size: 56 kDa,68 kDa

false

• WB

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Western blot - Anti-ATP6V1A antibody [EPR19270] (AB199326)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ATP6V1A antibody [EPR19270] (ab199326) at 1/2000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 20 µg

Lane 3:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 68 kDa

Observed band size: 68 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-ATP6V1A antibody [EPR19270] (AB199326)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 3 minutes; Lane 2 and 3 : 8 seconds.

All lanes:

Western blot - Anti-ATP6V1A antibody [EPR19270] (ab199326) at 1/2000 dilution

Lane 1:

Human fetal heart lysate at 10 µg

Lane 2:

Human fetal liver lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

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

false

• WB

Supplier Data

Western blot - Anti-ATP6V1A antibody [EPR19270] (AB199326)

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.

All lanes:

Western blot - Anti-ATP6V1A antibody [EPR19270] (ab199326) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Rat brain lysate at 10 µg

Lane 4:

Rat kidney lysate at 10 µg

Lane 5:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 6:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 7:

NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 68 kDa

Observed band size: 68 kDa

false