- PA5-19760 - Invitrogen Antibodies ACTR2 antibody

Cohen S, Kovari DT, Wei W, Keate R, Curtis JE, Nie S
Journal of molecular cell biology 2018 Oct 1;10(5):376-387

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Jurkat Whole Cell Lysate using Product # PA5-19760, Arp2 primary antibody at a dilution of 1 µg/mL (lane 1). Staining of MCF7 Whole Cell Lysate at a dilution of 1 µg/mL (lane 2). Blot treated with a secondary IR Dye680-conjugated Goat polyclonal anti-Rabbit antibody was used at a dilution of 1:10000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: KD of Arp2 was achieved by transfecting HeLa with Arp2 specific siRNAs (Silencer® select Product # s223082, s19644). Western blot analysis (Fig. a) was performed using whole cell extracts from the Arp2 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Arp2 Polyclonal Antibody (Product # PA5-19760, 1µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution). 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 Arp2. .

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-Arp2 Polyclonal Antibody (Product # PA5-19760) and a 42 kDa band corresponding to Arp2 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), RAW 264.7 (Lane 3), Jurkat (Lane 4), NIH/3T3 (Lane 5), Hep G2 (Lane 6), HEK-293 (Lane 7) and THP-1 (Lane 8) 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 µg/mL) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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)
Main image
Experimental details: Immunofluorescence analysis of Arp2 was performed using HeLa 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 Arp2 Polyclonal Antibody (Product # PA5-19760) at 1µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 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 nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..

PA5-19760