Anti-Aquaporin 2 antibody [EPR21080]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human kidney tissue staining CD34 with ab317588 at a 1/2000 dilution, ab199975 anti-Aquaporin 2 used at 1/4000 dilution and ab212197 anti-Renin used at a 1/4000 dilution.

Panel A : merged staining of anti-CD34 (green; Opal™520), anti-Aquaporin 2 (magenta; Opal™690) and anti-Renin (gray; Opal™570) on human kidney.

Panel B : anti-CD34 staining endothelium in human kidney.

Panel C : anti-Aquaporin 2 staining collecting tubules in human kidney.

Panel D : anti-Renin staining juxtaglomerular cells 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 ab317588, ab199975 and ab212197 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Aquaporin 2 with ab199975 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Luminal surface and tubular cytoplasmic staining on collecting duct of human kidney is observed (PMID : 15703994, PMID : 19546380). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human kidney tissue staining GLEPP1/PTPRO with ab322047 at a 1/2000 dilution, ab199975 anti-Aquaporin 2 used at 1/4000 dilution and ab212197 anti-Renin used at a 1/4000 dilution.

Panel A : merged staining of anti-GLEPP1/PTPRO (green; Opal™520), anti-Aquaporin 2 (magenta; Opal™690) and anti-Renin (gray; Opal™570) on human kidney.

Panel B : anti-GLEPP1/PTPRO staining glomerulus in human kidney.

Panel C : anti-Aquaporin 2 staining collecting tubules in human kidney.

Panel D : anti-Renin staining juxtaglomerular cells 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, ab199975 and ab212197 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

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Fluorescence multiplex immunohistochemical analysis of the human kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Hexokinase 1 (ab150423, green; Opal™690), anti-Angiotensin Converting Enzyme 1 (ab254222, gray; Opal™520) and anti-Aquaporin 2 (ab199975, red; Opal™570) on human kidney. Panel B : anti-Aquaporin 2 stained on collecting tubules. Panel C : anti-Angiotensin Converting Enzyme 1 stained on proximal tubules. Panel D : anti-Hexokinase 1 stained on distal tubules and collecting tubules. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The section was incubated in three rounds of staining : in the order of ab150423 at 1/250 dilution (4.224 μg/ml), ab254222 at 1/4000 dilution (0.141 μg/ml) and ab199975 at 1/4000 dilution (0.152 μg/ml) 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. DAPI (blue) was used as a nuclear counter stain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Aquaporin 2 with ab199975 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Luminal surface and tubular cytoplasmic staining on collecting duct of mouse kidney is observed (PMID : 15703994, PMID : 19546380). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Immunohistochemical analysis of 4% paraformaldehyde-fixed rat kidney tissue labeling Aquaporin 2 (green) with ab199975 at 1/3000 dilution followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Positive luminal membrane staining and tubular cytoplasmic staining on rat kidney is observed (PMID : 9688853; 15703994; 19546380). Heat-mediated antigen retrieval was performed using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).

Nuclear counterstain : DAPI (blue).
Permeabilization : 0.2% Triton X-100

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is AlexaFluor^®488 Goat anti-Rabbit secondary ab150077 at 1/1000 dilution.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human kidney tissue staining GLEPP1/PTPRO with ab322047 at a 1/2000 dilution, ab199975 anti-Aquaporin 2 used at 1/4000 dilution and ab212197 anti-Renin used at a 1/4000 dilution.

Panel A : merged staining of anti-GLEPP1/PTPRO (green; Opal™520), anti-Aquaporin 2 (magenta; Opal™690) and anti-Renin (gray; Opal™570) on human kidney.

Panel B : anti-GLEPP1/PTPRO staining glomerulus in human kidney.

Panel C : anti-Aquaporin 2 staining collecting tubules in human kidney.

Panel D : anti-Renin staining juxtaglomerular cells 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, ab199975 and ab212197 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

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining CD34 with ab317588 at a 1/5000 dilution, ab199975 anti-Aquaporin 2 used at 1/4000 dilution and ab212197 anti-Renin used at a 1/4000 dilution.

Panel A : merged staining of anti-CD34 (green; Opal™520), anti-Aquaporin 2 (magenta; Opal™690) and anti-Renin (gray; Opal™570) on human kidney.

Panel B : anti-CD34 staining endothelium in mouse kidney.

Panel C : anti-Aquaporin 2 staining collecting tubules in mouse kidney.

Panel D : anti-Renin staining juxtaglomerular cells in mouse kidney.

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

The section was incubated in three rounds of staining : in the order of ab317588, ab199975 and ab212197 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

Supplier Data

Multiplex immunohistochemistry - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining NADPH oxidase 4 with ab319055 at a 1 : 500 (1.032 ug/ml) dilution, ab199975 anti-Aquaporin 2 used at 1 : 4000 (0.143 ug/ml) dilution and ab254222 anti-Angiotensin Converting Enzyme 1 used at a 1 : 4000 (0.132 ug/ml) dilution.

Panel A : merged staining of anti-NADPH oxidase 4 (green; Opal™520), anti-Aquaporin 2 (magenta; Opal™690) and anti-Angiotensin Converting Enzyme 1 (yellow; Opal™570) on mouse kidney.
Panel B : anti-NADPH oxidase 4 staining distal tubules in mouse kidney.
Panel C : anti-Aquaporin 2 staining collecting tubules in mouse kidney.
Panel D : anti-Angiotensin Converting Enzyme 1 staining proximal tubules in mouse kidney.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab319055, ab199975 and ab254222 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.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat kidney tissue staining NADPH oxidase 4 with ab319055 at a 1 : 500 (1.032 ug/ml) dilution, ab199975 anti-Aquaporin 2 used at 1 : 4000 (0.143 ug/ml) dilution and ab254222 anti-Angiotensin Converting Enzyme 1 used at a 1 : 4000 (0.132 ug/ml) dilution.

Panel A : merged staining of anti-NADPH oxidase 4 (green; Opal™520), anti-Aquaporin 2 (magenta; Opal™690) and anti-Angiotensin Converting Enzyme 1 (yellow; Opal™570) on rat kidney.
Panel B : anti-NADPH oxidase 4 staining distal tubules in rat kidney.
Panel C : anti-Aquaporin 2 staining collecting tubules in rat kidney.
Panel D : anti-Angiotensin Converting Enzyme 1 staining proximal tubules in rat kidney.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab319055, ab199975 and ab254222 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat kidney tissue staining Renin with ab326049 at a 1 : 5000 (0.103 µg/ml) dilution ab317588 anti-CD34 used at 1 : 5000 (0.099 µg/ml) dilution and ab199975 anti-Aquaporin 2 used at a 1 : 4000 (0.152 µg/ml) dilution.

Panel A : merged staining of anti-Renin (green; Opal™520) anti-CD34 (magenta; Opal™690) and anti-Aquaporin 2 (gray; Opal™570) on rat kidney.
Panel B : anti-Renin staining juxtaglomerular cells in rat kidney.
Panel C : anti-CD34 staining endothelium in rat kidney.
Panel D : anti-Aquaporin 2 staining collecting tubules in rat kidney.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab326049 ab317588 and ab199975 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining Renin with ab326049 at a 1 : 5000 (0.103 µg/ml) dilution ab317588 anti-CD34 used at 1 : 5000 (0.099 µg/ml) dilution and ab199975 anti-Aquaporin 2 used at a 1 : 4000 (0.152 µg/ml) dilution.

Panel A : merged staining of anti-Renin (green; Opal™520) anti-CD34 (magenta; Opal™690) and anti-Aquaporin 2 (gray; Opal™570) on mouse kidney.
Panel B : anti-Renin staining juxtaglomerular cells in mouse kidney.
Panel C : anti-CD34 staining endothelium in mouse kidney.
Panel D : anti-Aquaporin 2 staining collecting tubules in mouse kidney.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab326049 ab317588 and ab199975 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.

• IHC-Fr

Unknown

Immunohistochemistry (Frozen sections) - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Immunohistochemical analysis of 4% paraformaldehyde-fixed mouse kidney tissue labeling Aquaporin 2 (green) with ab199975 at 1/3000 dilution followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution. Positive luminal membrane staining and tubular cytoplasmic staining on mouse kidney is observed (PMID : 9688853; 15703994; 19546380). Heat-mediated antigen retrieval was performed using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).

Nuclear counterstain : DAPI (blue).
Permeabilization : 0.2% Triton X-100

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is AlexaFluor^®488 Goat anti-Rabbit secondary ab150077 at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Aquaporin 2 with ab199975 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Luminal surface and tubular cytoplasmic staining on collecting duct of rat kidney is observed (PMID : 15703994, PMID : 19546380). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IP

Supplier Data

Immunoprecipitation - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

Aquaporin 2 was immunoprecipitated from 0.35 mg mouse kidney lysate with ab199975 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Mouse kidney lysate 10 μg (input).
Lane 2 : ab199975 IP in mouse kidney lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab199975 in mouse kidney lysate (-).

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

Exposure time : 10 seconds

We recommend customers optimize their lysis buffer and sample preparation for IP. This protein might aggregate during heating of samples.

The molecular mass is consistent with the literature (PMID 27405971; PMID 26109089).

All lanes:

Immunoprecipitation - Anti-Aquaporin 2 antibody [EPR21080] (ab199975)

Predicted band size: 28 kDa

false

• WB

Supplier Data

Western blot - Anti-Aquaporin 2 antibody [EPR21080] (AB199975)

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

Exposure time : 10 seconds.

We recommend customers optimize their lysis buffer and sample preparation for WB. This protein might aggregate during heating of samples.

The molecular mass is consistent with literature (PMID 27405971; PMID 26109089).

All lanes:

Western blot - Anti-Aquaporin 2 antibody [EPR21080] (ab199975) at 1/500 dilution

Lane 1:

Human fetal kidney lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

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: 28 kDa

false