Anti-Wnt3 antibody [EPR29462-511] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Wnt3 antibody [EPR29462-511] - BSA and Azide free (AB328049)

This data was developed using ab325601, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human small intestine tissue staining SLC6A19 with ab326561 at a 1 : 500 (0.996 µg/ml) dilution ab272692 anti-MUC2 used at 1 : 2000 (0.26 µg/ml) dilution and ab325601 anti-Wnt3 used at a 1 : 2000 (0.246 µg/ml) dilution.

Panel A : anti-SLC6A19 (green; Opal™520) anti-MUC2 (magenta; Opal™690) anti-Wnt3 (gray; Opal™570) on human small intestine.
Panel B : anti-SLC6A19 staining apical membrane of epithelium in human small intestine.
Panel C : anti-MUC2 staining goblet cells in human small intestine.
Panel D : anti-Wnt3 staining Paneth cells in human small intestine.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab326561 ab272692 and ab325601 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Wnt3 antibody [EPR29462-511] - BSA and Azide free (AB328049)

This data was developed using ab325601, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human ileum tissue staining Wnt3 with ab325601 at a 1/2000 (0.246 μg/ml) dilution ab272692 anti-MUC2 used at a 1/2000 (0.26 μg/ml) dilution and ab324982 anti-BTNL3 used at a 1/500 (0.106 μg/ml) dilution.

Panel A : anti-Wnt3 (green; Opal™520) anti-MUC2 (magenta; Opal™690) anti-btnl3 (gray; Opal™570) on human ileum.

Panel B : anti-Wnt3 staining Paneth cells in human ileum.

Panel C : anti-MUC2 staining goblet cells in human ileum.

Panel D : anti-BTNL3 staining brush border in human ileum.

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

The section was incubated in three rounds of staining : in the order of ab325601 ab272692 and ab324982 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3 antibody [EPR29462-511] - BSA and Azide free (AB328049)

This data was developed using ab325601, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human embryonic kidney epithelial cell) transfected with a human WNT3 expression vector containing a His Myc tag (B) HEK-293T transfected with a human WNT3A expression vector containing a His Myc tag and (C) HEK-293T transfected with an empty expression vector containing a His Myc tag labeling Wnt3 with ab325601 at 1/2000 (0.246 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a human WNT3 expression vector containing a His Myc tag; no staining on both (B) HEK-293T transfected with a human WNT3A expression vector containing a His Myc tag and (C) HEK-293T transfected with an empty expression vector containing a His Myc tag.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3 antibody [EPR29462-511] - BSA and Azide free (AB328049)

This data was developed using ab325601, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling Wnt3 with ab325601 at 1/2000 (0.246 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on human skeletal muscle.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3 antibody [EPR29462-511] - BSA and Azide free (AB328049)

This data was developed using ab325601, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Wnt3 with ab325601 at 1/2000 (0.246 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining on human spleen.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3 antibody [EPR29462-511] - BSA and Azide free (AB328049)

This data was developed using ab325601, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling Wnt3 with ab325601 at 1/2000 (0.246 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon carcinoma.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3 antibody [EPR29462-511] - BSA and Azide free (AB328049)

This data was developed using ab325601, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human ileum tissue labeling Wnt3 with ab325601 at 1/2000 (0.246 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Paneth cells of human ileum.

The primary antibody was incubated 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