Extended Data Fig. 3: Comparison of SNAP-tag2 und SNAPf-tag for labeling with different fluorescent substrates in live cells.
U2OS cell stably expressing mEGFP-SNAP-tag2 or mEGFP-SNAPf-tag were incubated with different a TMR, b CPY, c SiR, d MaP555 or e MaP618 substrates at [100 nM] or [500 nM] overnight or at [100 nM, 50 nM and 10 nM] for 1 h. Cells were washed and analyzed by flow cytometry (recorded n = 10000 cells). Analysis was done using FlowJo by gating for single cells and double positive labeling signal (as depicted in Supplementary Fig. 16) and the median of the fluorescent label/mEGFP was derived. Data are represented as median values ± mean s.d. of technical duplicates. SNAP-tag2 showed for most of the substrates a higher labeling ratio.
