Anti-Claudin 4 antibody [EPRR17575]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane and staining on epithelial cells of human colon 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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

Immunohistochemical analysis of paraffin-embedded human cholangiocarcinoma tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on human cholangiocarcinoma 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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on epithelial cells of human breast cancer 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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Weak membrane staining on biliary epithelium of human liver 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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on epithelial cells of human endometrium 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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling Claudin 4 with ab210796 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative staining on human hepatocellular carcinoma.

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

Claudin 4 was immunoprecipitated from 1mg of SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate with ab210796 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab210796 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : SW480 whole cell lysate 10μg (Input).

Lane 2 : ab210796 IP in SW480 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab210796 in SW480 whole cell lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-Claudin 4 antibody [EPRR17575] (ab210796)

Predicted band size: 22 kDa

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• WB

Supplier Data

Western blot - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

False colour image of Western blot : Anti-Claudin 4 antibody [EPRR17575] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab210796 was shown to bind specifically to Claudin 4. A band was observed at 17 kDa in wild-type MCF7 cell lysates with no signal observed at this size in CLDN4 knockout cell line ab274946 (knockout cell lysate ab275004). To generate this image, wild-type and CLDN4 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Claudin 4 antibody [EPRR17575] (ab210796) at 1/1000 dilution

Lane 1:

Wild-type MCF7 cell lysate at 20 µg

Lane 2:

Western blot - Human CLDN4 knockout MCF7 cell lysate (ab275004)

Lane 3:

PC-3 cell lysate at 20 µg

Lane 4:

U-2 OS cell lysate at 20 µg

Lane 5:

SW480 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 17 kDa

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• WB

Supplier Data

Western blot - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1/2/3/6 : 1 minute; Lane 4 : 30 seconds; Lane 5 : 3 minutes.

Lanes 1 - 2:

Western blot - Anti-Claudin 4 antibody [EPRR17575] (ab210796) at 1/2000 dilution

Lanes 3 - 6:

Western blot - Anti-Claudin 4 antibody [EPRR17575] (ab210796) at 1/1000 dilution

Lane 1:

LNCaP (Human prostate cancer cell line) whole cell lysate at 10 µg

Lane 2:

PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 3:

SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 5:

SK-OV-3 (Human ovarian cancer cell line) whole cell lysate at 10 µg

Lane 6:

NIH:OVCAR-3 (Human ovary adenocarcinoma cell line) whole cell lysate at 10 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

Predicted band size: 22 kDa

Observed band size: 18 kDa

false

• WB

Lab

Western blot - Anti-Claudin 4 antibody [EPRR17575] (AB210796)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The identity of the higher MW band at approximately 35 kDa is unknown.

All lanes:

Western blot - Anti-Claudin 4 antibody [EPRR17575] (ab210796) at 1/1000 dilution

All lanes:

4T1 (mouse mammary gland carcinoma 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 1/100000 dilution

Predicted band size: 22 kDa

Observed band size: 18 kDa

false

Exposure time: 3min