Anti-MUC2 antibody [EPR23479-47]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat colon tissue labeling MUC2 with ab272692 at 1/500 dilution followed by ab150077 AlexaFluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution (Green). Positive staining on rat colon is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 AlexaFluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human colon tissue staining GC-C with ab323804 at a 1/100 dilution, ab324525 anti-Peptide YY used at 1/2000 dilution and ab272692 anti-MUC2 used at a 1/2000 dilution.

Panel A : anti-GC-C (green; Opal™520), anti-Peptide YY (magenta; Opal™690) and anti-MUC2 (gray; Opal™570) on human colon.

Panel B : anti-GC-C staining apical membrane of epithelium in human colon.

Panel C : anti-Peptide YY staining enteroendocrine cells in human colon.

Panel D : anti-MUC2 staining goblet cells in human colon.

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

The section was incubated in three rounds of staining : in the order of ab323804, ab324525 and ab272692 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human small intestine staining SGLT1 with ab321787 at a 1/2000 dilution, ab272692 anti-MUS2 used at 1/2000 dilution and ab252935 anti-EPHB2 used at a 1/500 dilution.

Panel A : merged staining of anti-SGLT1 (green; Opal™520), anti-MUC2 (magenta; Opal™690) and anti-EPHB2 (gray; Opal™570) on human small intestine.

Panel B : anti-SGLT2 staining apical side of the villi in human small intestine.

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

Panel D : anti-EPHB2 staining stem 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 ab321787, ab272692 and ab252935 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-MUC2 antibody [EPR23479-47] (AB272692)

Fluorescence multiplex immunohistochemical analysis of the human colon (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Muc2 (ab272692, magenta; Opal™690), anti-Eph receptor B2 (ab252935, green; Opal™520) and anti-ErbB3 / HER3 (ab236220, red; Opal™570) on human colon. Panel B : anti-Muc2 stained on goblet cells. Panel C : anti-Eph receptor B2 stained on intestinal stem and progenitor cells. Panel D : anti-ErbB3 / HER3 stained on epithelial cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The section was incubated in three rounds of staining : in the order of ab272692 at 1/2000 dilution (0.252 μg/ml), ab252935 at 1/1000 dilution  (0.527 μg/ml), and ab236220 at 1/3000 dilution (0.75 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Fluorescence multiplex immunohistochemical analysis of the human colon (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Villin (ab245749, gray; Opal™690), anti-liver FABP (ab240401, green; Opal™520) and anti-MUC2 (ab272692, red; Opal™570) on human colon. Panel B : anti-liver FABP stained on enterocytes. Panel C : anti-Villin stained on apical border. Panel D : anti-MUC2 stained on goblet cells. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining : in the order of ab245749 (1/1000 dilution), ab240401 (1/8000 dilution), and ab272692 (1/5000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. 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. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human small intestine tissue staining MUC17 with ab316971 at 1/500 dilution, ab272692 anti-MUC2 used at 1/2000 dilution and ab323315 anti-REG1B used at a 1/5000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-MUC17 (green; Opal^™570), anti-MUC2 (magenta; Opal^™690) and anti-REG1B (grey; Opal^™520) on human small intestine.
Panel B : anti-MUC17 staining epithelium in human small intestine.
Panel C : anti-MUC2 staining goblet cells in human small intestine.
Panel D : anti-REG1B 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 ab316971, ab272692 and ab323315 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Fluorescence multiplex immunohistochemical analysis of the human jejunum (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Muc2 (ab272692, magenta; Opal™690), anti-Eph receptor B2 (ab252935, green; Opal™520) and anti-ErbB3 / HER3 (ab236220, red; Opal™570) on human jejunum. Panel B : anti-Muc2 stained on goblet cells. Panel C : anti-Eph receptor B2 stained on intestinal stem and progenitor cells. Panel D : anti-ErbB3 / HER3 stained on epithelial cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The section was incubated in three rounds of staining : in the order of ab272692 at 1/2000 dilution (0.252 μg/ml), ab252935 at 1/1000 dilution  (0.527 μg/ml), and ab236220 at 1/3000 dilution (0.75 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human ileum tissue staining BTNL3 with ab324982 at a 1/500 (0.106 μg/ml) dilution, ab272692 anti-MUC2 used at a 1/2000 (0.26 μg/ml) dilution and ab323315 anti-REG1B used at a 1/5000 (0.972 μg/ml) dilution.

Panel A : anti-BTNL3 (green; Opal™570), anti-MUC2 (magenta; Opal™690), anti-REG1B (gray; Opal™520) on human ileum.

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

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

Panel D : anti-REG1B staining Paneth cells 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 ab324982, ab272692 and ab323315 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-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling MUC2 with ab272692 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Postive staining on human colon (PMID : 28693267). The section was incubated with ab272692 for 10 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling MUC2 with ab272692 at 1/2000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Postive staining on human bladder cancer (PMID : 25197366). The section was incubated with ab272692 for 10 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse colon tissue staining GSDMC2 + GSDMC3 with ab229896 at a 1 : 2000 (0.25 ug/ml) dilution; SARM1 with ab309195 at 1 : 500 (0.964 ug/ml) dilution and MUC2 with ab272692 at 1 : 2000 (0.252 ug/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-SARM1 (green; Opal^™520), anti-MUC2 (gray; Opal^™570) and anti-GSDMC2/3 (magenta; Opal^™690)on mouse colon.
Panel B : anti-SARM1 staining myenteric nerve plexus in mouse colon.
Panel C : anti-MUC2 staining goblet cells in mouse colon.
Panel D : anti-GSDMC2/3 staining epithelium in mouse colon.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab309195, ab272692 and ab229896 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-MUC2 antibody [EPR23479-47] (AB272692)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse colon tissue staining Glycam1 with ab324912 at a 1/4000 ( 0.126 μg/ml) dilution, ab317313 anti-CD43 used at 1/500 dilution (1.052 μg/ml) and ab272692 anti-MUC2 used at 1/2000 dilution (0.26 μg/ml).

Panel A : anti-Glycam1 (green; Opal™520), anti-CD43 (magenta; Opal™690), anti-MUC2 (gray; Opal™570) on mouse colon.

Panel B : anti-Glycam1 staining HEV (highly endothelial venules) in mouse colon.

Panel C : anti-CD43 staining T lymphocytes in mouse colon.

Panel D : anti-MUC2 staining goblet cells in mouse colon.

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

The section was incubated in three rounds of staining : in the order of ab324912, ab237721 and ab315346 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse colon tissue staining GPA33 with ab324562 at a 1/100 dilution, ab324560 anti-ATAD2 used at 1/500 dilution and ab272692 anti-MUC2 used at a 1/2000 dilution.

Panel A : anti-GPA33 (green; Opal™690), anti-ATAD2 (magenta; Opal™520) and anti-MUC2 (gray; Opal™570) on mouse colon.

Panel B : anti-GPA33 staining membrane of epithelium in mouse colon.

Panel C : anti-ATAD2 staining nucleus of proliferating cells in mouse colon.

Panel D : anti-MUC2 staining goblet cells in mouse colon.

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

The section was incubated in three rounds of staining : in the order of ab324562, ab324560 and ab272692 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-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse colon tissue labeling MUC2 with ab272692 at 1/500 dilution followed by ab150077 AlexaFluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution (Green). Positive staining on mouse colon is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 AlexaFluor^® 488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling MUC2 with ab272692 at 1/2000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Postive staining on rat colon is observed. The section was incubated with ab272692 for 10 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling MUC2 with ab272692 at 1/2000 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Postive staining on mouse colon is observed. The section was incubated with ab272692 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections mouse small intestine tissue labelling Sodium/Hydrogen Exchanger 3 with ab307365 at 1 : 5000 dilution (B), MUC2 with ab272692 at 1 : 2000 dilution (C) Eph receptor B2 with ab252935 at 1 : 500 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Sodium/Hydrogen Exchanger 3 (green; Opal™520), anti-MUC2 (magenta; Opal™690) and anti-Eph receptor B2 (gray; Opal™570) on mouse small intestine.

Panel B : anti-Sodium/Hydrogen Exchanger 3 staining apical and luminal facing edges of surface cells in mouse small intestine.

Panel C : anti-MUC2 staining goblet cells in mouse small intestine.

Panel D : anti-Eph receptor B2 staining membrane of stem cells in mouse small intestine.

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

The section was incubated in three rounds of staining : in the order of ab307365, ab272692 and ab252935 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections rat colon tissue labelling Sodium/Hydrogen Exchanger 3 with ab307365 at 1 : 5000 dilution (B), MUC2 with ab272692 at 1 : 2000 dilution (C) Eph receptor B2 with ab252935 at 1 : 500 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Sodium/Hydrogen Exchanger 3 (green; Opal™520), anti-MUC2 (magenta; Opal™690) and anti-Eph receptor B2 (gray; Opal™570) on rat colon.

Panel B : anti-Sodium/Hydrogen Exchanger 3 staining apical and luminal facing edges of surface cells in rat colon.

Panel C : anti-MUC2 staining goblet cells in rat colon.

Panel D : anti-Eph receptor B2 staining membrane of stem cells in rat colon.

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

The section was incubated in three rounds of staining : in the order of ab307365, ab272692 and ab252935 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections mouse colon tissue labelling Sodium/Hydrogen Exchanger 3 with ab307365 at 1 : 5000 dilution (B), MUC2 with ab272692 at 1 : 2000 dilution (C) Eph receptor B2 with ab252935 at 1 : 500 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Sodium/Hydrogen Exchanger 3 (green; Opal™520), anti-MUC2 (magenta; Opal™690) and anti-Eph receptor B2 (gray; Opal™570) on mouse colon.

Panel B : anti-Sodium/Hydrogen Exchanger 3 staining apical and luminal facing edges of surface cells in mouse colon.

Panel C : anti-MUC2 staining goblet cells in mouse colon.

Panel D : anti-Eph receptor B2 staining membrane of stem cells in mouse colon.

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

The section was incubated in three rounds of staining : in the order of ab307365, ab272692 and ab252935 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse small intestine tissue staining Uridine Phosphorylase 1 with ab325391 at a 1/2000 (0.258 μg/ml) dilution, ab307365 anti-Sodium/Hydrogen Exchanger 3/NHE-3 used at a 1/5000 (0.126 μg/ml) dilution and ab272692 anti-MUC2 used at a 1/2000 (0.26 μg/ml) dilution.

Panel A : anti-Uridine Phosphorylase 1 (green; Opal™690), anti-Sodium/Hydrogen Exchanger 3/NHE-3 (magenta; Opal™520), anti-MUC2 (gray; Opal™570) on mouse small intestine.

Panel B : anti-Uridine Phosphorylase 1 staining cytoplasm and nucleus of epithelium in mouse small intestine.

Panel C : anti-Sodium/Hydrogen Exchanger 3/NHE-3 staining apical and luminal facing edges of surface cells in mouse small intestine.

Panel D : anti-MUC2 staining on goblet cells in mouse small intestine.

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

The section was incubated in three rounds of staining : in the order of ab325391, ab307365 and ab272692 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.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC2 antibody [EPR23479-47] (AB272692)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections rat small intestine tissue labelling Sodium/Hydrogen Exchanger 3 with ab307365 at 1 : 5000 dilution (B), MUC2 with ab272692 at 1 : 2000 dilution (C) Eph receptor B2 with ab252935 at 1 : 500 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Sodium/Hydrogen Exchanger 3 (green; Opal™520), anti-MUC2 (magenta; Opal™690) and anti-Eph receptor B2 (gray; Opal™570) on rat small intestine.

Panel B : anti-Sodium/Hydrogen Exchanger 3 staining apical and luminal facing edges of surface cells in rat small intestine.

Panel C : anti-MUC2 staining goblet cells in rat small intestine.

Panel D : anti-Eph receptor B2 staining membrane of stem cells in rat small intestine.

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

The section was incubated in three rounds of staining : in the order of ab307365, ab272692 and ab252935 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

• WB

Lab

Western blot - Anti-MUC2 antibody [EPR23479-47] (AB272692)

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

MUC2 is highly glycosylated.

The C-terminal fragment alpha-MUC2C2 (140-170 kDa) is observed. The molecular weight is consistent with what have been described in literature (PMID : 12582180; 16754877). Exposure time : 3 seconds.

All lanes:

Western blot - Anti-MUC2 antibody [EPR23479-47] (ab272692) at 1/1000 dilution

Lane 1:

Human colon cancer tissue lysate at 20 µg

Lane 2:

Mouse small intestine tissue lysate at 20 µg

Lane 3:

Rat small intestine tissue lysate at 20 µg

Lane 4:

Rat colon tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 540 kDa

Observed band size: 140-170 kDa

false