Western blot was performed using Anti-ZNF598 Recombinant Rabbit Monoclonal Antibody (Product # 703601) and a ~110 kDa band corresponding to ZNF598 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of HCT 116 (Lane 1) and HEK-293 (Lane 2) were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0302BOX). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:8000 dilution). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
For immunofluorescence analysis, HEK-293 cells were fixed and permeabilized for detection of endogenous ZNF598 using Anti-ZNF598 Recombinant Rabbit Monoclonal Antibody (Product # 703601, 1:100 dilution) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of ZNF598 protein (green), Panel b) is stained for nuclei (blue) using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of ZNF598. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supplier Page from Thermo Fisher Scientific for ZNF598 Antibody