Anti-Aquaporin 5 antibody [EPR27474-70]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lung tissue staining CD62L with ab325322 at a 1/1000 (0.509 μg/ml) dilution, ab305303 anti-Aquaporin-5 used at a 1/2000 (0.25 μg/ml) dilution and ab324911 anti-ATP-binding cassette sub-family A member 3 used at a 1/2000 (0.253 μg/ml) dilution.

Panel A : anti-CD62L (green; Opal™520), anti-Aquaporin-5 (magenta; Opal™690), anti-ATP-binding cassette sub-family A member 3 (gray; Opal™570) on mouse lung.

Panel B : anti-CD62L staining immune cells in mouse lung.

Panel C : anti-Aquaporin-5 staining type Ⅰ pneumocytes in mouse lung.

Panel D : anti-ATP-binding cassette sub-family A member 3 staining type Ⅱ pneumocytes in mouse lung.

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

The section was incubated in three rounds of staining : in the order of ab325322, ab305303 and ab324911 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Aquaporin 5 with ab305303 at 1/2000 (0.265 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Negative control : no staining on mouse kidney. The section was incubated with ab305303 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Aquaporin 5 with ab305303 at 1/2000 (0.265 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Negative control : no staining on rat kidney. The section was incubated with ab305303 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney (fresh) tissue labeling Aquaporin 5 with ab305303 at 1/100 (5.3 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). Negative control : confocal image showing no staining on rat kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab305303 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh) tissue labeling Aquaporin 5 with ab305303 at 1/100 (5.3 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). Negative control : confocal image showing no staining on mouse kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab305303 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Aquaporin 5 with ab305303 at 1/2000 (0.265 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on mouse lung. The section was incubated with ab305303 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lung tissue staining ATP-binding cassette sub-family A member 3 with ab324911 at a 1 : 2000 (0.253 ug/ml) dilution, ab322404 anti-PSAP used at 1 : 5000 (0.092 ug/ml) dilution and ab305303 anti-Aquaporin-5 used at a 1 : 2000 (0.265 ug/ml) dilution.

Panel A : merged staining of anti-ATP-binding cassette sub-family A member 3 (green; Opal™690), anti-PSAP (magenta; Opal™520) and anti-Aquaporin-5 (yellow; Opal™570) on mouse lung.
Panel B : anti-ATP-binding cassette sub-family A member 3 staining typeⅡ pneumocytes in mouse lung.
Panel C : ant-PSAP staining typeⅡ pneumocytes in mouse lung.
Panel D : ant-Aquaporin-5 staining typeⅠ pneumocytes in mouse lung.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324911, ab322404 and ab305303 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling Aquaporin 5 with ab305303 at 1/2000 (0.265 µg/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Positive staining on rat lung. The section was incubated with ab305303 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung (fresh) tissue labeling Aquaporin 5 with ab305303 at 1/100 (5.3 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). Confocal image showing positive staining on mouse lung. The nuclear counterstain was DAPI (Blue). The section was incubated with ab305303 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat lung (fresh) tissue labeling Aquaporin 5 with ab305303 at 1/100 (5.3 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). Confocal image showing positive staining on rat lung. The nuclear counterstain was DAPI (Blue). The section was incubated with ab305303 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Aquaporin 5 was immunoprecipitated from 0.35 mg Rat lung tissue lysate 10 μg with ab305303 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305303. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Rat lung tissue lysate 10 μg Lane 2 : ab305303 IP in Rat lung tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305303 in Rat lung tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 15 seconds Samples are non-boiled as boiling may cause protein aggregation.

All lanes:

Immunoprecipitation - Anti-Aquaporin 5 antibody [EPR27474-70] (ab305303) at 1/30 dilution

All lanes:

Rat lung tissue

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

Observed band size: 24 kDa

false

Exposure time: 15s

• IP

Supplier Data

Immunoprecipitation - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Aquaporin 5 was immunoprecipitated from 0.35 mg Mouse lung tissue lysate 10 μg with ab305303 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305303. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse lung tissue lysate 10 μg Lane 2 : ab305303 IP in Mouse lung tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305303 in Mouse lung tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 seconds Samples are non-boiled as boiling may cause protein aggregation.

All lanes:

Immunoprecipitation - Anti-Aquaporin 5 antibody [EPR27474-70] (ab305303) at 1/30 dilution

All lanes:

Mouse lung tissue

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

Observed band size: 24 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : kidney (PMID : 9767520). Exposure time : 48 seconds

All lanes:

Western blot - Anti-Aquaporin 5 antibody [EPR27474-70] (ab305303) at 1/1000 dilution

Lane 1:

Mouse lu tissue lysate prepared in RIPA lysis method at 20 µg

Lane 2:

Rat lu tissue lysate prepared in RIPA lysis method at 20 µg

Lane 3:

Mouse kidney tissue lysate prepared in RIPA lysis method at 20 µg

Lane 4:

Rat kidney tissue lysate prepared in RIPA lysis method 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 1/50000 dilution

Observed band size: 24 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-Aquaporin 5 antibody [EPR27474-70] (AB305303)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : cerebellum (PMID : 9767520), heart (PMID : 9767520). Exposure time : 70 seconds

All lanes:

Western blot - Anti-Aquaporin 5 antibody [EPR27474-70] (ab305303) at 1/1000 dilution

Lane 1:

Mouse salivary gland tissue lysate prepared in RIPA lysis method at 20 µg

Lane 2:

Mouse cerebellum tissue lysate prepared in RIPA lysis method at 20 µg

Lane 3:

Mouse heart tissue lysate prepared in RIPA lysis method 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 1/50000 dilution

Observed band size: 24 kDa

false

Exposure time: 70s