Transfected HEK293 cells with GFP-Cas9-SA fusion protein were stained with Anti-Cas9 from Staphylococcus Aureus Antibody. GFP is expressed in the transfected cells (green). GFP-Cas9 fusion protein is stained with Anti-Cas9 from Staphylococcus Aureus Antibody (1:1,000 | red). Only transfected cells are reactive with Anti-Cas9 from Staphylococcus Aureus Antibody, which appear in a yellow-orange color (merge). Nuclei are visualized in blue with Hoechst staining.
Immunofluorescent analysis of HEK293 cells transfected with a construct of green fluorescent protein (GFP) fused with the C-terminal sequence (aa. 803-1053) of S. aureus Cas9. Cells were stained with Anti-Cas9 Antibody [6F7] (A85319) in red, and this combined with the expressed GFP gives a green signal. The two signals overlap as expected. The nuclei of both transfected and untransfected cells are revealed by the DAPI DNA stain.
Figure 3: Western Blot - Anti-CRISPR-Cas9 Antibody [6F7] (A85319)
Western blot analysis of HEK293 cell lysates using Anti-Cas9 Antibody [6F7] (A85319), in green. The lanes contain samples of: [1] Protein standards, in red, [2] non-transfected cells, and [3] cells transfected with a GFP-CAS9 fusion construct, encoding the C-terminal (803-1053) amino acids of S. aureus CAS9. The strong band at about 53 kDa corresponds to the GFP-CAS9 fusion protein.
Figure 4: Western Blot - Anti-CRISPR-Cas9 Antibody [6F7] (A85319)
Western blot analysis of Anti-Cas9 from Staphylococcus Aureus Antibody (1:1,000). 1: HEK293 cells which overexpressed fusion protein containing GFP and c-terminus of Cas9 from S. aureus. 2: Non-transfected HEK293 cells. There is a strong clean band at about 53 kDa corresponding to GFP-Cas9 fusion protein, which is absent in non-transfected cells.
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