PA5-47441

antibody from Invitrogen Antibodies

Targeting: NECTIN3 CD113, CDw113, DKFZP566B0846, nectin-3, PPR3, PVRL3, PVRR3

Antibody data

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Validation data

Product number: PA5-47441 - Provider product page
Provider: Invitrogen Antibodies
Product name: Nectin 3 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant full-length protein
Description: In direct ELISAs, less than 65% cross-reactivity with recombinant mouse Nectin-3 is observed and less than 5% cross-reactivity with recombinant human (rh) Nectin-1, rhNectin-2, and rhNectin-4 is observed.
Concentration: 0.2 mg/mL

NECTIN3 protein structure - PA5-47441 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Nectin 3 was performed using 70% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Nectin 3 Goat Polyclonal Antibody (Product # PA5-47441) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11078) at a dilution of 1:2000 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). Panel d represents the merged image showing membrane and cell junctional localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: KD of Nectin 3 was achieved by transfecting Caco-2 cells with Nectin 3 specific siRNA (Silencer® select Product # S24804). Immunofluorescence analysis was performed on untransfected Caco-2 cells (panel a,d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with Nectin 3 specific siRNA (panel c,f). Cells were fixed, permeabilized, and labelled with Nectin 3 Goat Polyclonal Antibody (Product # PA5-47441, 5 µg/mL), followed by Rabbit anti-Goat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11078, 1:2000). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Reduction of specific signal was observed upon siRNA mediated KD (panel c,f) confirming specificity of the antibody to Nectin 3 (green). The images were captured at 60X magnification.