Anti-MUC17 antibody [EPR28482-85]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-MUC17 antibody [EPR28482-85] (AB316971)

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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MUC17 antibody [EPR28482-85] (AB316971)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed 0.1% TritonX-100 permeabilized AsPC-1 (human Pancreas epithelial cell) cells labelling MUC17 with ab316971 at 1/50 (10.2 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing cytoplasmic staining in AsPC-1 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : K-562 (PMID : 11855812)
Image was taken with a confocal microscope(Leica-Microsystems TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/mL) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MUC17 antibody [EPR28482-85] (AB316971)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized AsPC-1 (human Pancreas epithelial cell) / right and K-562 (human chronic myelogenous leukemia lymphoblast) / left cells labelling MUC17 with ab316971 at 1/500 dilution (0.1ug)/(Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : K-562

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC17 antibody [EPR28482-85] (AB316971)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling MUC17 with ab316971 at 1/500 (1.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human tonsil.
The section was incubated with ab316971 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC17 antibody [EPR28482-85] (AB316971)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling MUC17 with ab316971 at 1/500 (1.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human kidney.
The section was incubated with ab316971 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-MUC17 antibody [EPR28482-85] (AB316971)

Immunohistochemical analysis of paraffin-embedded Human jejunum tissue labeling MUC17 with ab316971 at 1/500 (1.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Membranous staining on epithelium of human jejunum.
The section was incubated with ab316971 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-MUC17 antibody [EPR28482-85] (AB316971)

Immunohistochemical analysis of paraffin-embedded Human stomach cancer tissue labeling MUC17 with ab316971 at 1/500 (1.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Membranous staining on epithelium of human stomach cancer (PMID : 31262330).
The section was incubated with ab316971 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-MUC17 antibody [EPR28482-85] (AB316971)

Immunohistochemical analysis of paraffin-embedded Human pancreas cancer tissue labeling MUC17 with ab316971 at 1/500 (1.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Membranous staining on epithelium of human pancreas cancer (PMID : 16737958).
The section was incubated with ab316971 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-MUC17 antibody [EPR28482-85] (AB316971)

Immunohistochemical analysis of paraffin-embedded (A) AsPC-1 (human Pancreas epithelial cell), (B) K-562 (human chronic myelogenous leukemia lymphoblast) tissue labeling MUC17 with ab316971 at 1/500 (1.02 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on panel (A) AsPC-1 cell pellet, no staining on panel (B) K-562 cell pellet.
The section was incubated with ab316971 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

• WB

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Western blot - Anti-MUC17 antibody [EPR28482-85] (AB316971)

Negative control : K-562, Hela (PMID : 11855812).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11855812, 16737958).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-MUC17 antibody [EPR28482-85] (ab316971) at 1/1000 dilution

Lane 1:

AsPC-1 (human Pancreas epithelial cell) whole cell lysate at 20 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 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 1/20000 dilution

Observed band size: 500 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-MUC17 antibody [EPR28482-85] (AB316971)

Negative control : kidney (PMID : 11855812).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-MUC17 antibody [EPR28482-85] (ab316971) at 1/1000 dilution

Lane 1:

Human small intestine tissue lysate at 20 µg

Lane 2:

Human kidney tissue lysate at 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 1/2000 dilution

Observed band size: 500 kDa,124 kDa

true

Exposure time: 180s