Table 2 Numerical values reporting on the association rate constant, k_on, and the dissociation rate constant, k_off, quantifying the interaction between different oligonucleotides and BsCspB obtained under dilute conditions and in crowded environments determined by kinetic stopped-flow fluorescence spectroscopy.

Ligand	Condition	k_on (10⁷/s/M)	k_off (/s)	k_off^eq (/s)
dT5	Dilute	34 ± 1	100 ± 10	260
dT6	Dilute	32 ± 1	n.d	18
dT7	Dilute	35 ± 4	n.d	4
dT5	100 g/L EG	25 ± 2	120 ± 10	n.d.
dT5	200 g/L EG	15 ± 1	160 ± 10	n.d.
dT5	300 g/L EG	10 ± 3	230 ± 10	510
dT5	100 g/L PEG1	23 ± 1	100 ± 10	n.d.
dT5	200 g/L PEG1	13 ± 1	100 ± 10	n.d.
dT5	300 g/L PEG1	7 ± 1	110 ± 5	270
dT5	100 g/L glucose	29 ± 1	40 ± 10	n.d.
dT5	200 g/L glucose	18 ± 1	40 ± 10	n.d.
dT5	300 g/L glucose	10 ± 3	40 ± 5	130
dT5	100 g/L Dex20	27 ± 2	70 ± 20	190
dT5	200 g/L Dex20	8 ± 1	85 ± 5	50
dT5	300 g/L PEG1	7 ± 1	110 ± 5	266
dT6	300 g/L PEG1	7.8 ± 0.1	10 ± 1	40
dT7	300 g/L PEG1	7.5 ± 0.1	3.1 ± 0.3	13

The value of k_off^eq has been calculated by applying k_off^eq = K_D*k_on (see Table 1 reporting on K_D values). The concentration of BsCspB was set to 60 nM in all experiments except in presence of 300 g/L EG (c^BsCspB = 300 nM). A 5- to 20-fold stoichiometric excess of ligand concerning BsCspB has been used in all kinetic stopped-flow fluorescence experiments which have been performed at T = 288 K. n.d.: not determined. Associated data are shown in Figs. 3A–D, S7A,B, and S8A–C.

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