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. Author manuscript; available in PMC: 2011 Nov 16.
Published in final edited form as: Biochemistry. 2010 Oct 25;49(45):9935–9941. doi: 10.1021/bi1014437

Figure 1.

Figure 1

Attachment of polyP to solid supports. (A) Dose response for covalently immobilizing polyP onto Amine Surface microplates. Varying polyPHMW concentrations were reacted overnight at 37°C in microplate wells in the absence (○) or presence (•) of 100 mM EDAC, after which unbound polyP was removed by washing with 2 M LiCl. Data are mean ± S.E.M. (n=3). (B) Influence of polymer length on coupling efficiency. PolyP preparations of varying mean polymer lengths were reacted overnight at 37°C with Sepabeads EC-HA and 100 mM EDAC with 1 mM CaCl2, after which unbound polyP was removed by washing with 2 M LiCl and 10 mM EDTA. PolyP concentrations (given in terms of phosphate) were adjusted to yield a constant 1 mM ends: 1 mM monophosphate and pyrophosphate; 1.5 mM triphosphate; 2.5 mM polyP5; 7 mM polyP14; 12.5 mM polyP25; 22.5 mM polyP45; 30 mM polyP60; and 65 mM polyP130. Controls plotted on the right include 50 mM monophosphate as well as reactions without EDAC. Data are expressed as mean percent recoveries of offered polyP (± S.E.M.; n=3 to 5). In both panels, bound polyP was quantified as monophosphate following acid hydrolysis.