Immunocytochemical analysis of CTCF in NIH-3T3 cells using a CTCF Monoclonal antibody (Product # MA5-32640) as seen in red. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Western blot was performed using Anti-CTCF Recombinant Rabbit Monoclonal Antibody (JM10-61) (Product # MA5-32640) and a 110 kDa band corresponding to CTCF and an uncharacterized band (*) at ~55kDa was observed across all the cell lines tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of Fig (a) HeLa (Lane 1), PC-12 (Lane 2), HEK-293 (Lane 3), NIH/3T3 (Lane 4), HCT 116 (Lane 5), Jurkat (Lane 6), RAW 264.7 (Lane 7), Hep G2 (Lane 8); Fig (b) MCF7 (Lane 1) and MCF7 treated with Trichostatin A (10uM for 24hr) (Lane 2) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 (1:500 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).