- PA5-28419 - Invitrogen Antibodies PPP2CA antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using PP2A alpha Polyclonal Antibody (Product # PA5-28419). Sample (50 µg of whole cell lysate). Lane A: mouse brain. 10% SDS PAGE. PP2A alpha Polyclonal Antibody (Product # PA5-28419) diluted at 1:10,000. 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: Western Blot using PP2A alpha Polyclonal Antibody (Product # PA5-28419). Sample (30 µg of whole cell lysate). Lane A: 293T. Lane B: Molt-4. 10% SDS PAGE. PP2A alpha Polyclonal Antibody (Product # PA5-28419) diluted at 1:1500. 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 PP2A alpha was achieved by transfecting PC-3 cells with PP2A alpha specific siRNAs (Silencer® select Product # s10958). Western blot analysis (Fig. a) was performed using whole cell extracts from the PP2A alpha knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-PP2A alpha Polyclonal Antibody (Product # PA5-28419, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25 µg/ml, 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 PP2A alpha.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-PP2A alpha Polyclonal Antibody (Product # PA5-28419) and 36 kDa band corresponding to PP2A alpha was observed across the cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HL-60 (Lane 1), A-431 (Lane2), PC-3 (Lane 3), HEL 92.1.7 (Lane 4), K-562 (Lane 5), A549 (Lane 6), HeLa (Lane 7), tissue extracts (30 µg lysate) of Mouse Brain (Lane 8) and Mouse Kidney (Lane 9) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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 analysis of PP2A alpha in paraformaldehyde-fixed HeLa cells using a PP2A alpha polyclonal antibody (Product # PA5-28419) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of PP2A alpha was performed using 70% confluent log phase PC-3 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 PP2A alpha Polyclonal Antibody (Product # PA5-28419) at 1:500 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034) 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, cytoskeletal 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: Immunohistochemical analysis of paraffin-embedded A549 xenograft, using PPP2CA (Product # PA5-28419) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunoprecipitation of PP2A alpha was performed in 293T whole cell extracts using 5 µg of PP2A alpha Polyclonal Antibody (Product # PA5-28419). Samples were transferred to a membrane and probed with PP2A alpha Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.

PA5-28419