Transfected HEK293 cells overexpressing a GFP-Cas9-SA fusion protein were stained with Anti-Cas9 from Staphylococcus Aureus Antibody. Cells which are transfected with GFP-Cas9 are bright green (left panel). Staining with Anti-Cas9 from Staphylococcus Aureus Antibody is shown in red (middle panel). In merged image (right panel), most HEK293 cells are not transfected so only the nucleus of these cells can be visualized with a blue DNA stain. Red antibody staining is only seen in cells which express GFP, as expected, and the superimposition of green and red results in an orange signal.
Immunofluorescent analysis of HEK293 cells transfected with a construct including the C-terminal 250 amino acids of S. aureus Cas9 fused to GFP, and then stained with Anti-Cas9 Antibody (A85318) in red. Transfected cells express the green fusion protein and bind the antibody in red, producing a yellow signal. Nuclear DNA in transfected and non-transfected cells is revealed with the blue DNA stain DAPI.
Figure 3: Western Blot - Anti-CRISPR-Cas9 Antibody (A85318)
Western blot analysis of HEK293 cell lysates using Anti-Cas9 Antibody (A85318), in green. The lanes contain samples of: [1] Protein standards, in red, [2] nontransfected cells and [3] cells transfected with a GFP-CAS9 fusion construct, encoding the C-terminal (803-1053) amino acids of S. aureus CAS9. The band at about 53 kDa corresponds to the GFP-CAS9 fusion protein.
Figure 4: Western Blot - Anti-CRISPR-Cas9 Antibody (A85318)
Western blot analysis of Anti-Cas9 from Staphylococcus Aureus Antibody(1:2,000). 1: HEK293 cells overexpressing a fusion protein containing GFP and the C-terminus of Cas9 from S. aureus. 2: Non-transfected HEK293 cells. The band at about 53 kDa corresponds to the GFP-Cas9 fusion protein and it is absent from non-transfected cells.
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