- PA5-27413 - Invitrogen Antibodies ARHGDIA antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of RhoGDI using 30 µg of IMR32 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a RhoGDI polyclonal antibody (Product # PA5-27413) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using RhoGDI Polyclonal Antibody (Product # PA5-27413). Sample (50 µg of whole cell lysate). Lane A: mouse brain. 12% SDS PAGE. RhoGDI Polyclonal Antibody (Product # PA5-27413) diluted at 1:10,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using RhoGDI Polyclonal Antibody (Product # PA5-27413). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with RhoGDI Polyclonal Antibody (Product # PA5-27413) diluted at 1:500. 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 RhoGDI was achieved by transfecting HeLa with RhoGDI specific siRNAs (Silencer® select Product # s698, s699). Western blot analysis (Fig. a) was performed using Whole Cell Extract-WCL from the RhoGDI knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with RhoGDI Polyclonal Antibody (Product # PA5-27413, 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 RhoGDI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-RhoGDI Polyclonal Antibody(Product # PA5-27413) and a 25kDa band corresponding to RhoGDI was observed across the cell lines tested. Whole Cell Extract-WCL (30 ug µg lysate) of HEK-293 (Lane 1), SH-SY5Y (Lane 2), U-937 (Lane 3), NIH:OVCAR-3 (Lane 4), COS-7 (Lane 5), NIH/3T3 (Lane 6), HeLa (Lane 7) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) 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: Immunofluorescence analysis of RhoGDI was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 45 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with RhoGDI Polyclonal Antibody (Product # PA5-27413) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), 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 dilution). Panel d represents the merged image showing Cytoplasmic and slight nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X with Oil immersion magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded HUH7 xenograft, using RhoGDI (Product # PA5-27413) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

PA5-27413