Western blot analysis was performed on whole cell extracts (30 µg lysate) of HT-1080 (Lane 1), MDA-MB-231 (Lane 2) and A549 (Lane 3). The blot was probed with Rabbit Anti-MMP14 Polyclonal Antibody (Product # PA5-16514, 2µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 65 kDa band corresponding to MMP14 was observed across the cell lines tested, an additional band of 160 kDa corresponding to the oligomer form of MMP-14 was also detected. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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 following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Western blot analysis was performed on whole cell extracts (30 µg lysate) of HT-1080 (Lane 1), MDA-MB-231 (Lane 2) and A549 (Lane 3). The blot was probed with Rabbit Anti-MMP14 Polyclonal Antibody (Product # PA5-16514, 2µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 65 kDa band corresponding to MMP14 was observed across the cell lines tested, an additional band of 160 kDa corresponding to the oligomer form of MMP-14 was also detected. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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 following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supplier Page from Thermo Fisher Scientific for MMP14 Antibody