- PA5-52198 - Invitrogen Antibodies WASL antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of N-WASP in human cell line RT-4 using a N-WASP Polyclonal Antibody (Product # PA5-52198).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of N-WASP in mouse cell line NIH-3T3 and rat cell line NBT-II using a N-WASP Polyclonal Antibody (Product # PA5-52198).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of N-WASP was achieved by transfecting MCF7 with N-WASP specific siRNAs (Silencer® select Product # S17133, S17132). Western blot analysis (Fig. a) was performed using Whole cell extracts from the N-WASP knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with N-WASP Polyclonal Antibody (Product # PA5-52198, 0.04 µg/mL) 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 N-WASP.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-N-WASP Polyclonal Antibody (Product # PA5-52198) and a ~60 kDa band corresponding to N-WASP was observed along with an uncharacterized band (*) at ~35 kDa across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of SK-BR-3 (Lane 1), SH-SY5Y (Lane 2), MCF7 (Lane 3), NIH/3T3 (Lane 4), PC-12 (Lane 5); and tissue extract Mouse Lung (Lane 6) and Mouse Cerebellum (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 (0.04 µg/mL) 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 staining of N-WASP in human cell line A-431 shows positivity in cytoplasm. Samples were probed using a N-WASP Polyclonal Antibody (Product # PA5-52198).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of N-WASP was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 5 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with N-WASP Polyclonal Antibody (Product # PA5-52198) at 20 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (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). Panel d represents the merged image showing Nucleus and cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60 magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of N-WASP in human cerebral cortex and skeletal muscle tissues using N-WASP Polyclonal Antibody (Product # PA5-52198). Corresponding WASL RNA-seq data are presented for the same tissues.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of N-WASP in human cerebral cortex using N-WASP Polyclonal Antibody (Product # PA5-52198) shows moderate to strong positivity in neuronal cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of N-WASP in human kidney using N-WASP Polyclonal Antibody (Product # PA5-52198) shows moderate to strong positivity in cells in tubules and glomeruli.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of N-WASP in human stomach using N-WASP Polyclonal Antibody (Product # PA5-52198) shows moderate to strong positivity in glandular cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical staining of N-WASP in human skeletal muscle using N-WASP Polyclonal Antibody (Product # PA5-52198) shows weak positivity in myocytes.

PA5-52198