- PA5-21405 - Invitrogen Antibodies ARPC2 antibody

Paingankar MS, Arankalle VA
Virus research 2015 Oct 2;208:98-109

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of ARPC2 using 30 µg of A) 293T and B) HeLa S3 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with an ARPC2 polyclonal antibody (Product # PA5-21405) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using ARPC2 Polyclonal Antibody (Product # PA5-21405). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with ARPC2 Polyclonal Antibody (Product # PA5-21405) diluted at 1:2,000. 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: ARPC2 Polyclonal Antibody detects ARPC2 protein by western blot analysis. A. 50 µg mouse kidney lysate/extract.10% SDS-PAGE. ARPC2 Polyclonal Antibody (Product # PA5-21405) dilution: 1:2,000. 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: ARPC2 Polyclonal Antibody detects ARPC2 protein by western blot analysis. A. 50 µg rat kidney lysate/extract.10% SDS-PAGE. ARPC2 Polyclonal Antibody (Product # PA5-21405) dilution: 1:5,000. 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: KD of ARPC2 was achieved by transfecting A-431 cells with ARPC2 specific siRNAs (Silencer® select Product # s19670, s19671). Western blot analysis (Fig. a) was performed using whole cell extracts from the ARPC2 KD cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with ARPC2 Polyclonal Antibody (Product # PA5-21405, 1:2500 dilution) and Rabbit anti-Goat IgG (H+L) Superclonal™ Secondary Antibody, HRP (Product # A27014, 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 ARPC2. .

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-ARPC2 Rabbit Polyclonal Antibody (Product # PA5-21405) and a 35kDa band corresponding to ARPC2 was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), U-87 MG (Lane 2), A-431 (Lane 3), Caco-2 (Lane 4), Jurkat (Lane 5), NIH/3T3 (lane 6), PC-12 (Lane 7), Mouse Kidney (Lane 8) and Rat 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 (1:2500 dilution) and detected by chemiluminescence Rabbit Anti-Goat IgG Secondary Antibody, HRP conjugate (Product # A27014, 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 ARPC2 in paraformaldehyde-fixed HeLa cells using an ARPC2 polyclonal antibody (Product # PA5-21405) at a 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of paraformaldehyde-fixed HeLa, using ARPC2 antibody (Product # PA5-21405) at 1:200 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of ARPC2 was performed using 70% confluent log phase A-431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ARPC2 Polyclonal Antibody (Product # PA5-21405) at 1:150 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Rabbit anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A-11078) 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 cytoplasmic and 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: Immunohistochemical analysis of paraffin-embedded OVCA xenograft, using ARPC2 (Product # PA5-21405) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

PA5-21405