- PA5-55012 - Invitrogen Antibodies DDX6 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of DDX6 in mouse cell line NIH-3T3 and rat cell line NBT-II using a DDX6 Polyclonal Antibody (Product # PA5-55012).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of DDX6 using DDX6 Polyclonal Antibody (Product # PA5-55012) (A) shows similar pattern to an independent DDX6 Polyclonal Antibody (B).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of DDX6 was achieved by transfecting HeLa with DDX6 specific siRNAs (Silencer® select Product # s4010, s4011). Western blot analysis (Fig. a) was performed using whole cell extracts from the DDX6 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DDX6 Polyclonal Antibody (Product # PA5-55012, 1:500 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ 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 DDX6. Uncharacterized band (*) at ~ 38 kDa was also observed in the samples tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using DDX6 Polyclonal Antibody (Product # PA5-55012) and a 54 kDa band corresponding to DDX6 was observed across all cell lines along with an uncharacterized band (*) at ~38 kDa in the human cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Daudi (Lane 2), Jurkat (Lane 3), Hep G2 (Lane 4), A-431 (Lane 5), NIH/3T3 (Lane 6) and PC-12 (Lane 7) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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: Immunofluorescent staining of DDX6 in human cell line U-251 MG shows positivity in cytoplasm & vesicles. Samples were probed using a DDX6 Polyclonal Antibody (Product # PA5-55012).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent staining of DDX6 in human cell line A-431 using a DDX6 Polyclonal Antibody (Product # PA5-55012) shows localization to cytoplasmic bodies.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of DDX6 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with DDX6 Polyclonal Antibody (Product # PA5-55012) 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 1:2000 dilution 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 cytoplasmic 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)
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Experimental details: Immunohistochemical staining of DDX6 in human tonsil using a DDX6 Polyclonal Antibody (Product # PA5-55012) shows strong cytoplasmic positivity in germinal and non-germinal center cells.

PA5-55012