Western blot analysis was performed on (Fig 1) whole cell extracts (30 µg lysate) of HeLa (Lane 1), HT-29 (Lane 2), SK-OV-3 (Lane 3), MCF7 (Lane 4), PC-3 (Lane 5), K-562 (Lane 6) and HCT 116 (Lane 7). Likewise Western blot analysis was performed on (Fig 2) 10 uL HeLa conditioned media (Lane 1) and 10 uL MDA-MB-231 conditioned media (Lane 2). The blots were probed with Anti-TIMP-1 Receptor Mouse Monoclonal Antibody (Product # MS608PABX, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 23 kDa band corresponding to TIMP-1 was observed across the cell lines and conditioned media tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).