Anti-MUC13 antibody [EPR21901]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC13 antibody [EPR21901] (AB235450)

Immunohistochemical analysis of paraffin-embedded human colorectal cancer tissue labeling MUC13 with ab235450 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive cytoplasmic staining in human colorectal cancer (PMID : 17360025) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC13 antibody [EPR21901] (AB235450)

Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling MUC13 with ab235450 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive cytoplasmic staining in human gastric cancer (PMID : 15904467) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MUC13 antibody [EPR21901] (AB235450)

Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol permeabilized LoVo (human colorectal adenocarcinoma cell line) cells labeling MUC13 with ab235450 at 1/40 (red) compared with a Isotype control details (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-MUC13 antibody [EPR21901] (AB235450)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized LoVo (human colorectal adenocarcinoma cell line) cells labeling MUC13 with ab235450 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in the LoVo cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Use PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC13 antibody [EPR21901] (AB235450)

Immunohistochemical analysis of paraffin-embedded human colorectal cancer tissue labeling MUC13 with ab235450 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the apical surface and in the cytoplasm in human colorectal cancer (PMID : 17360025) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MUC13 antibody [EPR21901] (AB235450)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling MUC13 with ab235450 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in epithelial cells of the normal human colon (PMID : 17360025) is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-MUC13 antibody [EPR21901] (AB235450)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT-29 (human colorectal adenocarcinoma cell line) cells labeling MUC13 with ab235450 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in the HT-29 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Use PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• WB

Unknown

Western blot - Anti-MUC13 antibody [EPR21901] (AB235450)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-3 and 5 : 3 minutes, Lane 4 : 26 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID : 15904467). Lane 3 was developed using a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-MUC13 antibody [EPR21901] (ab235450) at 1/1000 dilution

Lane 1:

HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

SW480 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 3:

HCT 116 (human colorectal carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Human ovary cancer lysate at 10 µg

Lane 5:

LoVo (human colorectal adenocarcinoma cell line) whole cell lysate at 10 µg

Secondary

Lanes 1 - 3:

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

Lanes 4 - 5:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Predicted band size: 55 kDa

Observed band size: 120 kDa,35 kDa,80 kDa

false

• WB

Lab

Western blot - Anti-MUC13 antibody [EPR21901] (AB235450)

Western blot : Anti-MUC13 antibody [EPR21901] ab235450 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 100-150 kDa in Wild-type A549 cell lysates with no signal observed at this size in MUC13 knockout A549 cell line. Two close non-specific bands were detected near to MUC13. We have not invesitgated the identity of these bands. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween 20 (TBS-T) before incubation with primary antibodies overnight at 4 C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-MUC13 antibody [EPR21901] (ab235450) at 1/100 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

MUC13 knockout A549 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 100-150 kDa

Observed band size: 100-150 kDa

false