14-2237-80

antibody from Invitrogen Antibodies

Targeting: CDH3 CDHP, PCAD

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

14-2237-80

Provider

Invitrogen Antibodies

Product name

P-Cadherin Monoclonal Antibody (CSTEM29), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

This monoclonal antibody CSTEM29 reacts with human P-Cadherin, also known as Cadherin-3.

Antibody clone number

CSTEM29

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of reduced murine C2C12 myoblast cells, H9 human ESCs, or human iPSCs using 10 µg/mL of Anti-Human P-Cadherin Purified antibody. Bands were visualized using Anti-Mouse IgG HRP.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-P-Cadherin Monoclonal Antibody (CSTEM29), eBioscience™ (Product # 14-2237-82) and 100kDa and 80kDa bands corresponding to Cadherin-3 was observed across all the tested cell lines and tissues, except HEL 92.1.7 and Mouse Skeletal Muscle. Whole cell extracts (40 µg lysate) of NIH:OVCAR-3 (Lane 1), U-2 OS (Lane 2), HEL 92.1.7 (Lane 3), Caco-2 (Lane 4), Mouse Ovary (Lane 5), Mouse Placenta (Lane 6), Mouse Heart (Lane 7), Mouse Skeletal Muscle (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (10 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Cadherin-3 was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.01% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with P-Cadherin Monoclonal Antibody (CSTEM29), eBioscience™ (Product # 14-2237-82) at 5µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cell junction localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Staining of a mixture of human iPSC and the C2C12 murine cell line with Anti-Human TRA-1-60 PerCP-eFluor 710 (Product # 46-8863-82) and 0.25 µg Mouse IgG2a K Isotype Control Purified (Product # 14-4724-82) (left) or 0.25 µg of Anti-Human P-Cadherin Purified (right) followed by F (ab')2 Anti-Mouse IgG PE (Product # 12-4010-82).