The Availability of a Recombinant Anti-SNAP Antibody in VHH Format Amplifies the Application Flexibility of SNAP-Tagged Proteins

Figure 4

Double staining of SNAP-tagged Twist2 obtained by coupling specific antibodies and benzylguanine-fluorochromes. HeLa cells were transiently transfected using 20 𝜇g/plate of plasmid for the expression of the SNAP-Twist2 protein. Its subcellular localization was identified: (a) and (e) by the in vivo addition of the permeable SNAP-Cell TMR Star fluorochrome (dilution 1 : 200); (b) by means of the purified recombinant anti-SNAP antibody 3G1B fused to the rabbit Fc (5 𝜇g/mL); (f) by means of the cell culture supernatant containing the recombinant anti-Twist2 VHH antibody 2C12 (dilution 1 : 2); (d) and (h) are the merged images of (a)+(b) and (e)+(f), respectively; (c) and (g) correspond to the nuclear DAPI staining; (i), (j), and (k) are nontransfected cells (controls) treated with the indicated antibodies. HeLa cells transfected with SNAP-Epsin8 were stained with SNAP-Cell TMR Star fluorochrome (dilution 1 : 200) to rule out a SNAP-dependent nuclear staining (l).

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