Anti-SLC12A3 antibody [EPR27106-48]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC12A3 antibody [EPR27106-48] (AB316191)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling SLC12A3 with ab316191 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on distal tubules of human kidney (PMID : 33328404).
The section was incubated with ab316191 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-SLC12A3 antibody [EPR27106-48] (AB316191)

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 ab316191 anti-SLC12A3 used at a 1 : 100 (5.08 ug/ml) dilution.

Panel A : merged staining of anti-PTPRP (magenta; Opal™690), anti-SLC5A2 (green; Opal™520) and anti-SLC12A3 (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-SLC12A3 staining distal 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 ab316191 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-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC12A3 antibody [EPR27106-48] (AB316191)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling SLC12A3 with ab316191 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on distal tubules of mouse kidney (PMID : 26337786).
The section was incubated with ab316191 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 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-SLC12A3 antibody [EPR27106-48] (AB316191)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling SLC12A3 with ab316191 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on distal tubules of rat kidney (PMID : 21865292).
The section was incubated with ab316191 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-SLC12A3 antibody [EPR27106-48] (AB316191)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining GLEPP1/PTPRO with ab322047 at a 1 : 2000 (0.261 ug/ml) dilution, ab316191 anti-SLC12A3 used at 1 : 2000 (0.254 ug/ml) dilution and ab254222 anti-Angiotensin Converting Enzyme 1 used at a 1 : 4000 (0.141 ug/ml) dilution.

Panel A : merged staining of anti-PTPRO (magenta; Opal™690), anti-SCL12A3 (green; Opal™520) and anti-Angiotensin Converting Enzyme 1 (gray; Opal™570) on mouse kidney.
Panel B : anti-PTPRO staining glomerulus in mouse kidney.
Panel C : anti-SCL12A3 staining distal 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 ab322047, ab316191 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.

• WB

Supplier Data

Western blot - Anti-SLC12A3 antibody [EPR27106-48] (AB316191)

Negative control : lung (PMID : 35591852), liver (PMID : 35591852).

Samples are non-boiled as boiling may cause protein aggregation.

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

All lanes:

Western blot - Anti-SLC12A3 antibody [EPR27106-48] (ab316191) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg with 5% NFDM/TBST

Lane 2:

Mouse liver tissue lysate at 20 µg with 5% NFDM/TBST

Lane 3:

Mouse lung tissue lysate at 20 µg with 5% NFDM/TBST

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 1/2000 dilution

Observed band size: 150 kDa,36 kDa

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SLC12A3 antibody [EPR27106-48] (AB316191)

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling SLC12A3 with ab316191 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on rat liver.
The section was incubated with ab316191 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 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-SLC12A3 antibody [EPR27106-48] (AB316191)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling SLC12A3 with ab316191 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human cerebrum (PMID : 8812482).
The section was incubated with ab316191 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 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-SLC12A3 antibody [EPR27106-48] (AB316191)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling SLC12A3 with ab316191 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human liver (PMID : 8812482).
The section was incubated with ab316191 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 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-SLC12A3 antibody [EPR27106-48] (AB316191)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling SLC12A3 with ab316191 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on rat cerebrum.
The section was incubated with ab316191 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 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-SLC12A3 antibody [EPR27106-48] (AB316191)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling SLC12A3 with ab316191 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse cerebrum.
The section was incubated with ab316191 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 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-SLC12A3 antibody [EPR27106-48] (AB316191)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SLC12A3 with ab316191 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse liver.
The section was incubated with ab316191 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Dot

Supplier Data

Dot Blot - Anti-SLC12A3 antibody [EPR27106-48] (AB316191)

Dot blot analysis of SLC12A3 using ab316191 at 1 : 1000 (0.508 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

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

This antibody does not cross-react with human SLC12A1 and human SLC12A2.

All lanes:

Dot Blot - Anti-SLC12A3 antibody [EPR27106-48] (ab316191) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a SLC12A3 expression vector containing a His-tag, whole cell lysate at 10 µg

Lane 3:

293T cells transfected with a SLC12A1 expression vector containing a His-tag, whole cell lysate at 10 µg

Lane 4:

293T cells transfected with a SLC12A2 expression vector containing a His-tag, whole cell lysate at 10 µg

Secondary

All lanes:

Dot 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

false

Exposure time: 169s