Fig. 3: Parameter space for covalent tetrazine labeling of receptor mutant K97B at the cell surface.
From: Insights into receptor structure and dynamics at the surface of living cells
a Single-cell average of click efficiency (CE) and receptor occupancy as a function of single-cell expression levels based on eGFP. The CE of K97B measured under standard labeling conditions is indicated by the gray shading (19% ± 3%, mean ± SD). The range of observed expression levels is discontinuous (arrows). The average CE and occupancy in this experiment (n = 119 cells) is indicated by dashed lines. b CE as a function of Alexa568-tet1 concentration (5 min incubation time); fitting follows a sublinear two-state saturation function. Markers and error bars represent mean ± SD of 20 cells. c CE as a function of incubation time (0.2 and 1.6 µM Alexa568-tet1); fitting follows a double exponential function. Markers and error bars represent mean ± SD of 50 (5 min) and 20 cells (else) in case of 0.2 µM or 20 (2 min) and 40 cells (else) in case of 1.6 µM Alexa568-tet1. d Influence of temperature on CE (1.6 µM Alexa568-tet1) after 5 and 30 min incubation time. Box-and-whisker plots indicate first and third quartile (box), median (horizontal line), and 1.5 times the interquartile range (whiskers) of 40 (30 min, 4 °C) or 20 cells (else). All experiments were performed once. Source data are provided as a Source Data file.
