Anti-Aquaporin 3 antibody [EPR28053-11]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell) (Left panel) compared to A431 (human epidermoid carcinoma epithelial cell) (Right panel) cells labelling Aquaporin 3 with ab307969 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• mIHC

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Multiplex immunohistochemistry - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human skin tissue labelling Aquaporin 3 with ab307969 at 1/5000 dilution (0.094 µg/mL) (B), Loricrin with ab176322 at 1/3000 dilution (0.031 μg/ml) (C) and Filaggrin with ab221155 at 1/2000 dilution (0.523 μg/ml) (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Panel A : merged staining of anti-Aquaporin 3 (magenta; Opal™690), anti-Loricrin (green; Opal™520) and anti-Filaggrin (red; Opal™570) on human skin. Panel B : anti-Aquaporin 3 stained on keratinocytes from the stratum basale to the stratum granulosum of the epidermis. Panel C : anti-Loricrin stained on stratum corneum. Panel D : anti-Filaggrin stained on the granular to stratum corneum transition. The section was incubated in three rounds of staining : in the order of ab307969, ab176322, and ab221155 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Aquaporin 3 with ab307969 at 1/5000 dilution (0.094 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Negative control : no staining on human cerebrum. The section was incubated with ab307969 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Aquaporin 3 with ab307969 at 1/5000 dilution (0.094 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human kidney. The section was incubated with ab307969 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Immunohistochemical analysis of paraffin-embedded human skin tissue labeling Aquaporin 3 with ab307969 at 1/5000 dilution (0.094 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human skin. The section was incubated with ab307969 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human skin staining Collagen VII with ab309143 at a 1/100 dilution, ab307969 anti-Aquaporin 3 used at 1/5000 dilution and ab221155 anti-FLG used at a 1/1000 dilution.

Panel A : merged staining of anti-Collagen VII (green; Opal™520), anti-Aquaporin 3 (magenta; Opal™690) and anti-FLG (gray; Opal™570) on human skin.

Panel B : anti-Collagen VII staining basement membrane in human skin.

Panel C : anti-Aquaporin 3 staining squamous epithelium in human skin.

Panel D : anti-FLG staining maturing squamous cells in human skin.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab309143, ab307969 and ab221155 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

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Multiplex immunohistochemistry - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human kidney tissue staining GLEPP1/PTPRO with ab322047 at a 1 : 2000 (0.261 ug/ml) dilution, ab320086 anti-SLC5A2 used at 1 : 1000 (0.508 ug/ml) dilution and ab307969 anti-Aquaporin 3 used at a 1 : 5000 (0.0941 ug/ml) dilution.

Panel A : merged staining of anti-PTPRP (magenta; Opal™690), anti-SLC5A2 (green; Opal™520) and anti-Aquaporin 3 (gray; Opal™570) on human kidney.
Panel B : anti-PTPRP staining glomerulus in human kidney.
Panel C : anti-SLC5A2 staining proximal tubules in human kidney.
Panel D : anti-Aquaporin 3 staining collecting tubules in human kidney.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab322047, ab320086 and ab307969 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

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Immunoprecipitation - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Aquaporin 3 was immunoprecipitated from 0.35 mg A431 (human epidermoid carcinoma epithelial cell) whole cell lysate with ab307969 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307969 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : A431 (human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 2 : ab307969 IP in A431 (human epidermoid carcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307969 in A431 (human epidermoid carcinoma epithelial cell) whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Aquaporin 3 antibody [EPR28053-11] (ab307969) at 1/30 dilution

All lanes:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 31 kDa

Observed band size: 31 kDa

false

• WB

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Western blot - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : brain(PMID : 10737773) In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds Negative control : brain(PMID : 10737773)

All lanes:

Western blot - Anti-Aquaporin 3 antibody [EPR28053-11] (ab307969)

Lane 1:

Human kidney tissue lysate at 20 mg/mL

Lane 2:

Human brain tissue lysate at 20 mg/mL

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 31 kDa

Observed band size: 35 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Aquaporin 3 antibody [EPR28053-11] (AB307969)

Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Aquaporin 3 antibody [EPR28053-11] (ab307969) at 1/1000 dilution

All lanes:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 31 kDa

Observed band size: 35 kDa

false

Exposure time: 180s