eIF3d Antibody PA5-97731 from Thermo Fisher Scientific

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WB

Western blot was performed using Anti-ACTN4 Polyclonal Antibody (Product # PA5-97731) and a 110 kDa band corresponding to Alpha-actinin-4 was observed across all the cell lines and tissue except LNCaP which is reported to be be a low expressing cell line. Whole cell extracts (30 µg lysate) of DU 145 (Lane 1), PC-3 (Lane 2), LNCaP (Lane 3), HeLa (Lane 4), A549 (Lane 5), A-431 (Lane 6), MCF7 (Lane 7) and PANC-1 (Lane 8) and tissue extract (30 µg lysate) of Mouse Skeletal Muscle (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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 (1:1000 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 Alpha-actinin-4 was performed using 70% confluent log phase PC-3 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 45 minutes at room temperature. The cells were labeled with ACTN4 Polyclonal Antibody (Product # PA5-97731) 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). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents LNCaP cells having low expression of GSTP1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

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Thermo Fisher Scientific for
eIF3d Antibody

Thermo Fisher Scientific's eIF3d Antibody is a Rabbit Polyclonal antibody. This antibody has been shown to work in applications such as: EIA, Immunoassay, ELISA, Immunocytochemistry, Immunohistochemistry, Immunohistochemistry - fixed, and Western Blot. The eIF3d Antibody was generated using EIF3D as the antigen and it reacts with Homo Sapiens, Human, and Rat.

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ICC/IF

eIF3d Antibody from Thermo Fisher Scientific

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eIF3d Antibody from Thermo Fisher Scientific

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