4.1.99.13 - 693832, 703212, 716329 4.1.99.13 (6-4) photoproduct DNA photolyase activity is detected in Crotalus atrox fibroblast. Activity is considerably enhanced when a UV-damaged DNA affinity column is used for purification. However, the activity is unstable and it is lost during purification or upon storage at -20° or -70°C for 2-3 months 682264 4.1.99.13 by fractionation of crude cell extracts with Heparin agarose and UV DNA affinity column chromatography 682086 4.1.99.13 by using a glutathione sepharose column and a Hi Trap Q column. Concentrated fusion protein is cleaved with thrombin from bovine plasma by incubation overnight at 4°C. For chromophore determination, the eluate from the glutathione sepharose column is purified through a Q sepharose column, omitting gel filtration procedure, and concentrated by ultrafiltration 682048 4.1.99.13 by using a glutathione-sepharose column and UV-irradiated DNA affinity column, fusion protein is cleaved with thrombin 682154 4.1.99.13 by using affinity chromatography and UV-irradiated DNA attached beads 682855 4.1.99.13 by using glutathione-sepharose columns 679920, 680605 4.1.99.13 fusion protein is applied to amylose column. At this point the protein is above 90% pure. Further purification can be obtained by applying the eluted material to a 10 ml heparin-agarose column. Maltose-binding protein is removed by treatment with factor Xa protease 681765 4.1.99.13 fusion protein is purified through a 20-ml amylose column and through heparin-agarose column 680604 4.1.99.13 NF-1 is isolated from nuclear extracts prepared from HeLa cell culture using a heparin column, an anion exchange Fractogel EMD TMAE-650 (S) column and a FPLC MonoS HR5/5 column 682153