Anti-Aquaporin 3 antibody

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody (AB125219)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Aquaporin 3 with ab125219 at 2 ug/ml. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C.

The section was blocked with 10% goat serum then incubated with ab125219 overnight at 4°C.

The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Aquaporin 3 antibody (AB125219)

Immunocytochemistry/ Immunofluorescence analysis of A431 (human epidermoid carcinoma cell line) cells labeling Aquaporin 3 with ab125219 at 5 μg/mL. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- AQP3 Antibody, ab125219, overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1 : 500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody (AB125219)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling Aquaporin 3 with ab125219 at 2 ug/ml. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C.

The section was blocked with 10% goat serum then incubated with ab125219 overnight at 4°C.

The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Aquaporin 3 antibody (AB125219)

Overlay histogram showing RT4 cells stained with ab125219 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with ab125219 (1µg/1x10⁶ cells) for 30 min at 20°C. DyLight^®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody (AB125219)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Aquaporin 3 with ab125219 at 2 ug/ml. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C.

The section was blocked with 10% goat serum then incubated with ab125219 overnight at 4°C.

The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody (AB125219)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Aquaporin 3 with ab125219 at 2 ug/ml. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C.

The section was blocked with 10% goat serum then incubated with ab125219 overnight at 4°C.

The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.

Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution).

• WB

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Western blot - Anti-Aquaporin 3 antibody (AB125219)

Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours.

After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. The membrane was then blocked with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Aquaporin 3 antigen affinity purified polyclonal antibody at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1 : 5000 for 1.5 hour at room temperature.

All lanes:

Western blot - Anti-Aquaporin 3 antibody (ab125219) at 0.5 µg/mL

Lane 1:

HeLa whole cell lysates at 30 µg

Lane 2:

Rat kidney tissue lysate at 30 µg

Lane 3:

Mouse kidney tissue lysate at 30 µg

Secondary

All lanes:

Goat anti-rabbit IgG-HRP at 1/5000 dilution

Observed band size: 61 kDa

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