Immunocytochemical analysis of DKK1 in NCCIT cells using a DKK1 Monoclonal antibody (Product # MA5-32229) as seen in green. 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-DKK1 Recombinant Rabbit Monoclonal Antibody (SC06-86) (Product # MA5-32229) and a 28-40 kDa band corresponding to DKK1 was observed in HCT 116 treated with Protein transport inhibitor (1X for 4hrs) but not in U-937 cells which is reported to low or negative for DKK1 expression. Additional uncharacterized bands (*) were observed across the cell lines. Whole-cell extracts (30 µg lysate) of HCT 116 (Lane 1), HCT 116 treated with Protein transport inhibitor (1X for 4hrs) (Lane 2), U-937 (Lane 3), U-937 treated with Protein transport inhibitor (1X for 4hrs) (Lane 4) 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 # A27036,1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).