Anti-P cadherin antibody [56/P-Cadherin]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-P cadherin antibody [56/P-Cadherin] (AB307740)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling P cadherin with ab307740 at 1/100 dilution (8.94 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Enhanced staining on human breast cancer (PMID : 18001487, PMID : 28084338). The section was incubated with ab307740 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is 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-P cadherin antibody [56/P-Cadherin] (AB307740)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling P cadherin with ab307740 at 1/100 dilution (8.94 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human breast (PMID : 18001487). The section was incubated with ab307740 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-P cadherin antibody [56/P-Cadherin] (AB307740)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1%Triton X-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling P cadherin with ab307740 at 1/100 dilution (8.94 ug/ml), followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing membranous staining in A431 cell line, no staining was observed in MCF7 cell line. Negative control : MCF7 (PMID : 10545506). Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) (Red). The nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

• WB

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Western blot - Anti-P cadherin antibody [56/P-Cadherin] (AB307740)

Blocking buffer : 5% NFDM/TBST. Exposure time : 136 seconds.

All lanes:

Western blot - Anti-P cadherin antibody [56/P-Cadherin] (ab307740) at 1/1000 dilution

Lane 1:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 120 kDa

false

Exposure time: 136s

• WB

Supplier Data

Western blot - Anti-P cadherin antibody [56/P-Cadherin] (AB307740)

Negative control : MCF7(PMID : 10545506). The molecular weight observed is consistent with what has been described in the literature (PMID : 2702654, 15548699) In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 26 seconds.

All lanes:

Western blot - Anti-P cadherin antibody [56/P-Cadherin] (ab307740) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 120 kDa

false

Exposure time: 26s