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Table 1 TAT-Cre recombinase in vitro stability in the presence of metal ions

From: Monodispersity of recombinant Cre recombinase correlates with its effectiveness in vivo

 

DLS

(RH in nm)

AI

(280em/340em nm)

Control

3.98 - monodispersed

0.02

1 mM NiCl2 at 4°C

polydispersed

8.5

10 mM NiCl2 at 4°C

polydispersed

9.2

1 mM NiCl2 at 20°C

-

precipitated

1 mM NiCl2 at 30°C

-

precipitated

0.1 mM NiCl2 at 4°C

polydispersed

3.1

0.01 mM NiCl2 at 4°C

4.15 - monodispersed

0.4

1 mM NiCl2 + 5 mM EDTA

polydispersed

3.3

1 mM NiCl2 + 10 mM EDTA

polydispersed

2.2

1 mM NiCl2 + 200 mM TA

3.92 - monodispersed

0.02

  1. Enzyme monodispersity was analyzed by dynamic light scattering (DLS) and fluorimetric aggregation index (AI). Purified recombinant protein samples (50 μg/mL) were incubated 30 min at different temperatures in the presence of increasing NiCl2 concentrations and of the chelators EDTA and tartaric acid (TA). DLS: the hydrodynamic radius (RH) of the molecules in solution is expressed in nm; the AI is the ratio between the light scattering at 280 nm and the emission at 340 nm. Experiments have been performed in triplicate.