- PA5-17471 - Invitrogen Antibodies DVL2 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Dvl2 in total cell extracts COS and Mv 1 Lu cells using Dvl2 polyclonal antibody (Product # PA5-17471).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Dvl2 in total cell extracts from NIH/3T3 cells, untreated or treated with conditioned medium containing Wnt3a for 3 hours, and also treated with or without lambda phosphatase, using Dvl2 polyclonal antibody (Product # PA5-17471).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of DVL2 was achieved by transfecting U-87 MG with DVL2 specific siRNAs (Silencer® select Product # s4398, s4397). Western blot analysis (Fig. a) was performed using whole cell extracts from the DVL2 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DVL2 Monoclonal Antibody (OTI7C7) (Product # PA5-17471, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to DVL2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-DVL2 Polyclonal Antibody (Product # PA5-17471) and a band at 90 kDa corresponding to DVL2 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of HCT 116 (Lane 1), HT-29 (Lane 2), Raji (Lane 3), U-87 MG (Lane 4), Jeko-1 (Lane 5), NIH/3T3 (Lane 6), MDA-MB-231 (Lane 7) and A549 (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12% % Bis-Tris gel (Product # NP0321BOX). 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of DVL2 was performed using 70% confluent log phase HCT 116 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 DVL2 Polyclonal Antibody (Product # PA5-17471) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

PA5-17471