- PA5-34696 - Invitrogen Antibodies PAK1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using PAK1 Polyclonal Antibody (Product # PA5-34696). Sample (30 µg of whole cell lysate). Lane A: A431. Lane B: HeLa. 7.5% SDS PAGE. PAK1 Polyclonal Antibody (Product # PA5-34696) diluted at 1:5,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PAK1 Polyclonal Antibody detects PAK1 protein by western blot analysis. Whole cell extracts (30 µg) was separated by 7.5% SDS-PAGE, and the membrane was blotted with PAK1 Polyclonal Antibody (Product # PA5-34696) diluted at 1:1,000. 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: PAK1 Polyclonal Antibody detects PAK1 protein by western blot analysis. A. 30 µg BCL-1 whole cell lysate/extract.7.5% SDS-PAGE. PAK1 Polyclonal Antibody (Product # PA5-34696) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: PAK1 Polyclonal Antibody detects PAK1 protein by western blot analysis. A. 30 µg PC-12 whole cell lysate/extract.7.5% SDS-PAGE. PAK1 Polyclonal Antibody (Product # PA5-34696) dilution: 1:1,000. 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: Knockdown of PAK1 was achieved by transfecting Caco-2 with PAK1 specific siRNAs (Silencer® select Product # S10020, S10021). Western blot analysis (Fig. A) was performed using Whole cell extracts from the PAK1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with PAK1 Polyclonal Antibody (Product # PA5-34696, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). Densitometric analysis of this western blot is shown in histogram (Fig. B). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to PAK1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-PAK1 Polyclonal Antibody (Product # PA5-34696) and a 60kDa band corresponding to PAK1 was observed across all the tested cell lines and tissues. Whole cell extracts (30 µg lysate) of Caco-2 (Lane 1), HEK-293 (Lane 2), Mouse Brain (Lane 3), Rat Brain (Lane 4) 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 (1:2000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of PAK1 showing staining in the cytoplasm of A431 cells. A431 cells were fixed in ice-cold MeOH for 5 min and stained using a PAK1 polyclonal antibody (Product # PA5-34696) diluted at 1:500. Blue: Hoechst 33343 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: PAK1 Polyclonal Antibody detects PAK1 protein at cytoplasm by immunofluorescent analysis. Sample: A431 cells were fixed in ice-cold MeOH for 5 min. Green: PAK1 protein stained by PAK1 Polyclonal Antibody (Product # PA5-34696) diluted at 1:500. Blue: Hoechst 33343 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of PAK1 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 45 minutes at room temperature. The cells were labeled with PAK1 Polyclonal Antibody (Product # PA5-34696) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (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, 1:300). Panel d represents the merged image showing a predominant nuclear 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: PAK1 Polyclonal Antibody detects PAK1 protein on human hepatoma by immunohistochemical analysis. Sample: Paraffin-embedded hepatoma. PAK1 Polyclonal Antibody (Product # PA5-34696) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

PA5-34696