Anti-Aquaporin 4 antibody [EPR24281-65]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling Aquaporin-4 with ab259318 at 1/2000 (0.299 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Strong membrane and cytoplasm staining on human cerebrum. (PMID : 7559426, 19170255) The section was incubated with ab259318 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval 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 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling Aquaporin-4 with ab259318 at 1/2000 (0.299 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human skeletal muscle. The section was incubated with ab259318 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval 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 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling Aquaporin-4 with ab259318 at 1/2000 (0.299 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human liver. (PMID : 19170255) The section was incubated with ab259318 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval 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 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling Aquaporin-4 with ab259318 at 1/2000 (0.299 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human kidney. (PMID : 19170255) The section was incubated with ab259318 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney (fresh) tissue labeling Aquaporin-4 with ab259318 at 1/500 (1.196 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Cytoplasmic and membranous staining on rat kidney (PMID : 17442782) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh) tissue labeling Aquaporin-4 with ab259318 at 1/500 (1.196 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse cerebrum (PMID : 21990350) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Rat primary neural/glia cell cells labelling Aquaporin-4 with ab259318 at 1/500 dilution (0.1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse testis (fresh) tissue labeling Aquaporin-4 with ab259318 at 1/500 (1.196 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Negative control : No staining on mouse testis testis (PMID : 16840793) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling Aquaporin-4 with ab259318 at 1/5000 (0.299 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab259318 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of 4% PFA-fixed 0.2% Triton X-100 permeabilized frozen Rat cerebrum (fresh) tissue labeling Aquaporin-4 with ab259318 at 1/500 (1.196 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on rat cerebrum (PMID : 21990350) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling Aquaporin-4 with ab259318 at 1/200 (2.99 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in rat astrocyte cells. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection is observed. ab10062 Anti-GFAP mouse monoclonal antibody was used as counterstain at 1/50 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Aquaporin-4 with ab259318 at 1/200 (2.99 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in mouse astrocyte cells. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection is observed. ab10062 Anti-GFAP mouse monoclonal antibody was used as counterstain at 1/50 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded (A) Skeletal muscle tissue from wild-type C57BL/6JGpt mice (B) Skeletal muscle tissue from AQP4 knockout mice labelling Aquaporin-4 with ab259318 at 1/5000 followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counterstained with Hematoxylin. Antigen retrieval was heat mediated with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Positive staining on (A) Skeletal muscle tissue from wild-type C57BL/6JGpt mice and no staining on (B) Skeletal muscle tissue from AQP4 knockout mice. The section was incubated with ab259318 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Aqp4-KO homozygous mice (Strain ID : T011901).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling Aquaporin-4 with ab259318 at 1/5000 (0.299 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse testis. The section was incubated with ab259318 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval 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 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling Aquaporin-4 with ab259318 at 1/5000 (0.299 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum. The section was incubated with ab259318 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse primary neural/glia cell cells labelling Aquaporin-4 with ab259318 at 1/500 dilution (0.1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse kidney (fresh) tissue labeling Aquaporin-4 with ab259318 at 1/500 (1.196 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Cytoplasmic and membranous staining on mouse kidney (PMID : 17442782) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat testis (fresh) tissue labeling Aquaporin-4 with ab259318 at 1/500 (1.196 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Negative control : No staining on rat testis testis (PMID : 16840793) is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded Mouse retina tissue labelling Aquaporin-4 with ab259318 at 1/5000 (0.299 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse retina. (PMID : 17525153) The section was incubated with ab259318 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 a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Immunohistochemical analysis of paraffin-embedded (A) Brain tissue from wild-type C57BL/6JGpt mice (B) Brain tissue from AQP4 knockout mice labelling Aquaporin-4 with ab259318 at 1/5000 followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Counterstained with Hematoxylin.  Antigen retrieval was heat mediated with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins Positive staining on (A) Brain tissue from wild-type C57BL/6JGpt mice and no staining on (B) Brain tissue from AQP4 knockout mice. The section was incubated with ab259318 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Aqp4-KO homozygous mice (Strain ID : T011901).

• WB

Lab

Western blot - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The molecular weight is consistent with what has been described in the literature. Bands observed at 52 kDa may represent the glycosylated form of AQP4 and the lower band may be the nonglycosylated form (PMID : 26786101, 8594871).

The expression profile observed in the human liver is consistent with the literature (PMID : 19170255).

Lysate were made unboil. We recommend not to boil the samples after lysis to get desired WB results.

All lanes:

Western blot - Anti-Aquaporin 4 antibody [EPR24281-65] (ab259318) at 1/1000 dilution

Lane 1:

Human skeletal muscle tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human testis tissue 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 1/100000 dilution

Predicted band size: 35 kDa

Observed band size: 28 kDa,52 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The molecular weight is consistent with what has been described in the literature. Bands observed at 100 kDa relate to a tetramer of AQP4 (PMID : 18334967, 11742978)

The expression profile observed in mouse testis is consistent with the literature (PMID : 16840793, 23712968).

Lysate were made unboil. We recommend not to boil the samples after lysis to get desired WB results.

All lanes:

Western blot - Anti-Aquaporin 4 antibody [EPR24281-65] (ab259318) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 3:

Mouse stomach tissue lysate at 20 µg

Lane 4:

Mouse testis tissue 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 1/100000 dilution

Predicted band size: 35 kDa

Observed band size: 100 kDa,28 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

In Western Blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) used at 1/200000 dilution.

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

Boiling leads to the formation of aggregates of membrane proteins, leading to unsuccessful migration in the gel. We recommend to to run unboiled lysate samples when testing for Aqp4 expression by western blot.

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Aqp4-KO homozygous mice (Strain ID : T011901).

All lanes:

Western blot - Anti-Aquaporin 4 antibody [EPR24281-65] (ab259318) at 1/1000 dilution

Lane 1:

Wild-type mouse brain tissue lysate (female)

Lanes 2 - 3:

AQP4 knockout brain tissue lysate (female)

Lane 4:

Wild-type mouse kidney tissue lysate (female)

Lanes 5 - 6:

AQP4 knockout kidney tissue lysate (female)

Lane 7:

Wild-type mouse skeletal muscle tissue lysate (female)

Lanes 8 - 9:

AQP4 knockout skeletal muscle tissue lysate (female)

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

Observed band size: 28 kDa,52 kDa

false

Exposure time: 4s

• WB

Lab

Western blot - Anti-Aquaporin 4 antibody [EPR24281-65] (AB259318)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The molecular weight is consistent with what has been described in the literature (PMID : 18334967, 11742978).

The expression profile observed in rat testis is consistent with the literature (PMID : 16840793, 23712968).

Lysate were made unboil. We recommend not to boil the samples after lysis to get desired WB results.

All lanes:

Western blot - Anti-Aquaporin 4 antibody [EPR24281-65] (ab259318) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat skeletal muscle tissue lysate at 20 µg

Lane 3:

Rat stomach tissue lysate at 20 µg

Lane 4:

Rat testis tissue 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 1/100000 dilution

Predicted band size: 35 kDa

Observed band size: 28 kDa

false

Exposure time: 3min