- PA5-34725 - Invitrogen Antibodies DBN1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of Drebrin was performed by separating 50 µg of Various tissue extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a Drebrin Polyclonal Antibody (Product # PA5-34725) at a dilution of 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using Drebrin Polyclonal Antibody (Product # PA5-34725). Sample (30 µg of whole cell lysate). Lane A: IMR32 . 7.5% SDS PAGE. Drebrin Polyclonal Antibody (Product # PA5-34725) diluted at 1:5,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Anti-Drebrin Polyclonal Antibody was achieved by transfecting HEK-293 cells with Drebrin specific siRNAs (Silencer® select Product # s3950, s3951). Western blot analysis (Fig. a) was performed using whole cell extracts from the Drebrin knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Drebrin Polyclonal Antibody, (Product # PA5-34725, 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 Drebrin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using anti-Drebrin Polyclonal Antibody (Product # PA5-34725) and a 80 kDa band corresponding to Drebrin was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), Jurkat (Lane 2), SH-SY5Y (Lane 3), PC-12 (Lane 4), tissue extracts of Mouse Brain (Lane 5) and Rat Brain (Lane 6) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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 using 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 Drebrin was performed using 70% confluent log phase HEK-293 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 10 minutes at room temperature. The cells were labeled with Drebrin Polyclonal Antibody (Product # PA5-34725) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A27034, 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 membrane and 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: Drebrin Polyclonal Antibody detects Drebrin protein at cytosol on rat hind brain by immunohistochemical analysis. Sample: Paraffin-embedded rat hind brain. Drebrin Polyclonal Antibody (Product # PA5-34725) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

PA5-34725