Histones from calf thymus

For separation of proteins in native agarose gels, recombinant VEGF (Life Technologies, Carlsbad, California), histones from calf thymus (Sigma-Aldrich, Steinheim, Germany) or catalase from bovine liver (Serva, Heidelberg, Germany) were each incubated with polySia (colominic acid from E. coli equal to polySia in mammals; GERBU, Heidelberg, Germany) in 50 mM Tris buffer for 1 h at 30 °C with agitation. The samples were loaded onto a 2% agarose gel (peqGOLD Universal Agarose, peqLab, Erlangen, Germany) and separated using 19.2 mM glycine in 25 mM Tris/HCl (pH 8.5) buffer. The electrophoresis was run at 80 V (constant voltage) for 3.5 h [32 (link)]. Thereafter, the gel was fixed in 45% methanol containing 7.5% acetic acid (v/v) overnight. Roti-blue (Roth, Karlsruhe, Germany) was used as Coomassie blue staining-dye according to manufacturer’s instructions.

Histones from calf thymus, bovine serum albumin, N-isopropylacrylamide (NIPAm), sodium dodecyl sulfate (SDS), fluoresceinamine isomer, and poly(ethylene glycol) methyl ether methacrylate (average M_n 500, 1500, and 4000) were purchased from Sigma Aldrich. N,N’-methylenebisacrylamide (BIS) was from Fluka; N-t-butylacrylamide (TBAm) was from ACROS ORGANICS. [2,3-14C]-Acrylamide was purchased from GE Healthcare UK Ltd. (Buckinghamshire, UK). Histone H1, H2A, H2B, H3.3 and H4 (Human, Recombinant) were purchased from New England Biolabs, Inc., (MA, USA). Sulfo-Cy5 NHS ester was purchased from Lumiprobe (FL, USA). 5-(and 6)-carboxytetramethylrhodamine, succinimidyl ester (NHS-Rhodamine) was purchased from Thermo Fisher Scientific, Inc.