CRISPR-Cas9 mediated genome editing ofAxl (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR932417_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of AXL was performed by loading 30 µg of U-87 MG Cas9 (Lane 1), and U-87 MG Cas9 cells transduced with Axl Lentiviral sgRNA (Lane 2) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ Novex™ 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 Anti-Axl Recombinant Rabbit Monoclonal Antibody (15H4L2) (Product # 702896) using 1:5,000 dilution and Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036 1:5,000 dilution).Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A loss of signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toAxl (Fig (b)).
CRISPR-Cas9 mediated genome editing ofAxl (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR932417_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of AXL was performed by loading 30 µg of U-87 MG Cas9 (Lane 1), and U-87 MG Cas9 cells transduced with Axl Lentiviral sgRNA (Lane 2) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ Novex™ 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 Anti-Axl Recombinant Rabbit Monoclonal Antibody (15H4L2) (Product # 702896) using 1:5,000 dilution and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036 1:5,000 dilution).Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A loss of signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toAxl (Fig (b)).
Supplier Page from Thermo Fisher Scientific for Axl Antibody