NEDD8 Antibody PA5-17476 from Thermo Fisher Scientific

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WB

Western blot was performed using anti-NEDD8 Polyclonal Antibody (Product # PA5-17476) and a 9 kDa band corresponding to NEDD8 was observed across all the cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HEK-293 (Lane 2), Jurkat (Lane 3), PC-12 (Lane 4), THP-1 (Lane 5), K-562 (Lane 6), HL-60 (Lane 7), Raw 264.7 (Lane 8), tissue extracts of Rat testis (Lane 9), Mouse brain (Lane 10) and Mouse colon (Lane 11) were electrophoresed Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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 (Heavy Chain), 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).
ICC/IF

Immunofluorescence analysis of NEDD8 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 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with NEDD8 Polyclonal Antibody (Product # PA5-17476) at 1:100 dilution 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) 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 localization to the cytoplasm and nucleus. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
NEDD8 Antibody

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NEDD8 Antibody

The NEDD8 Antibody from Thermo Fisher Scientific is a Rabbit Polyclonal antibody to NEDD8, and NEDD8 ubiquitin like modifier. This antibody recognizes Homo Sapiens, Human, Mouse, Rat, and Non-Human Primate antigen. The NEDD8 Antibody has been shown to work in the following applications: Immunocytochemistry, Immunohistochemistry, Immunohistochemistry - fixed, Immunoprecipitation, and Western Blot.

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NEDD8 Antibody

NEDD8 Antibody from Thermo Fisher Scientific

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NEDD8 Antibody from Thermo Fisher Scientific

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