Anti-SGLT2 antibody [EPR27112-7]

• IHC

Supplier Data

Immunohistochemistry - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Immunohistochemical analysis of paraffin-embedded transfected HEK-293T cells labeling SGLT2 with ab306558 at 1/5000 (0.104 μg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on HEK-293T cells transfected with a mouse SGLT2 expression vector (Panel A); No staining on HEK-293T cells transfected with a mouse Slc5a1 expression vector (Panel B); No staining on HEK-293T cells transfected with a mouse Slc5a3 expression vector (Panel C); No staining on HEK-293T cells transfected with an empty vector (Panel D). The section was incubated with ab306558 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat pancreas (fresh) tissue labeling SGLT2 with ab306558 at 1/1000 (0.52 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Negative control : confocal image showing no staining in rat pancreas (PMID : 33815487). The nuclear counterstain was DAPI (Blue). The section was incubated with ab306558 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 (2 ug/mL) dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse pancreas (fresh) tissue labeling SGLT2 with ab306558 at 1/1000 (0.52 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Negative control : confocal image showing no staining in mouse pancreas (PMID : 33815487). The nuclear counterstain was DAPI (Blue). The section was incubated with ab306558 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 (2 ug/mL) dilution.

• IHC

Supplier Data

Immunohistochemistry - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue to label SGLT2 with ab306558 at 1/5000 (0.104 μg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit). Negative control : no staining on mouse pancreas. The section was incubated with ab306558 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 min.

• IHC

Lab

Immunohistochemistry - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse kidney tissue staining PAR1/Thrombin Receptor with ab322457 at a 1/500 dilution, ab306558 anti-SGLT2 used at 1/5000 dilution and ab212197 anti-Renin used at a 1/4000 dilution.

Panel A : merged staining of anti-PAR1/Thrombin Receptor (green; Opal™520), anti-SGLT2 (magenta; Opal™690) and anti-Renin (gray; Opal™570) on mouse kidney.

Panel B : anti-PAR1/Thrombin Receptor staining endothelium in mouse kidney.

Panel C : anti-SGLT2 staining the brush border of proximal 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 ab322457, ab306558 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

Supplier Data

Immunohistochemistry - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling SGLT2 with ab306558 at 1/5000 (0.104 μg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit). Positive staining on the brush border of mouse proximal tubules. The section was incubated with ab306558 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Primary diluent was used instead of primary antibody, followed by anti-Mouse IgG1 antibody (ab125913). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 min.

• IHC

Supplier Data

Immunohistochemistry - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling SGLT2 with ab306558 at 1/5000 (0.104 μg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit). Positive staining on the brush border of rat proximal tubules. The section was incubated with ab306558 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Primary diluent was used instead of primary antibody, followed by anti-Mouse IgG1 antibody (ab125913). Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 min.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse kidney (fresh) tissue labeling SGLT2 with ab306558 at 1/1000 (0.52 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing positive staining in the proximal tubules of mouse kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab306558 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 (2 ug/mL) dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat kidney (fresh) tissue labeling SGLT2 with ab306558 at 1/1000 (0.52 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing positive staining in the proximal tubules of rat kidney. The nuclear counterstain was DAPI (Blue). The section was incubated with ab306558 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) preadsorbedat 1/1000 (2 ug/mL) dilution.

• WB

Supplier Data

Western blot - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Blocking / Diluting buffer and concentration : 5% NFDM/TBST Negative control : mouse pancreas (PMID : 30416006). Samples are non-boiled as boiling may cause protein aggregation.

All lanes:

Western blot - Anti-SGLT2 antibody [EPR27112-7] (ab306558) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate 20 μg

Lane 2:

Mouse pancreas tissue lysate 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

Observed band size: 73 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-SGLT2 antibody [EPR27112-7] (AB306558)

Samples are non-boiled as boiling may cause protein aggregation. Blocking / Diluting buffer and concentration : 5% NFDM/TBST. Exposure time : 15 seconds.

All lanes:

Western blot - Anti-SGLT2 antibody [EPR27112-7] (ab306558) at 1/1000 dilution

Lane 1:

Mouse kidney membrane fraction 20 μg

Lane 2:

Mouse kidney cytoplasmic fraction 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

Observed band size: 73 kDa

false

Exposure time: 15s