Western blot analysis was performed on Whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), U-87 MG (Lane 2), SH-SY5Y (Lane 3), NTERA-2 (Lane 4), MDA-MB-231 (Lane 5), T-47D (Lane 6), U-2-OS (Lane 7) and A-431 (Lane 8). The blots were probed with Anti-NOTCH3 Recombinant Rabbit Monoclonal Antibody (Product # 701975, 1-2 µg/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 110 kDa (cleaved fragment) band corresponding to NOTCH3 was observed across the cell lines tested. 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 wet transfer method. 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 MCF-7 (Lane 1), U-87 MG (Lane 2), SH-SY5Y (Lane 3), NTERA-2 (Lane 4), MDA-MB-231 (Lane 5), T-47D (Lane 6), U-2-OS (Lane 7) and A-431 (Lane 8). The blots were probed with Anti-NOTCH3 Recombinant Rabbit Monoclonal Antibody (Product # 701975, 1-2 µg/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 110 kDa (cleaved fragment) band corresponding to NOTCH3 was observed across the cell lines tested. 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 wet transfer method. 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 NOTCH3 Antibody