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. Author manuscript; available in PMC: 2013 Oct 17.
Published in final edited form as: Bioconjug Chem. 2012 Sep 25;23(10):2007–2013. doi: 10.1021/bc300052u

Figure 6.

Figure 6

(A) TL reactions of proteins 21–23 with biotin tetrazine 24b to generate 26–28 respectively. All reactions included 10-fold excess 24b and were conducted at room temperature (3 h) then overnight (4 °C) in PBS, pH 7.4. (B and C) Reaction products 26–28 were first incubated at the indicated concentrations (5 µg/mL data shown in B) with FR(+) HeLa cells and then analyzed by flow cytometry as described above. The upper limit of biotinylation defined as MFI exhibited by 19. (B) Biotinylation efficiency expressed as 100 × (MFI26, 27, or 28 / MFI19). (C) Maximum fluorescence signal defined as MFI of 19 at 10 µg/mL. All MFI data shown in C was normalized to MFI of 19 at 10 µg/mL.