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. 2002 May 15;30(10):2232–2243. doi: 10.1093/nar/30.10.2232

Figure 3.

Figure 3

Resolution of delta helicase and hnRNP L on FPLC TK119/RNA–Sepharose chromatography. (A) Helicase activity was assayed in the presence of ATP, reaction products were resolved on 8% non-denaturing PAGE gels, and products were detected by phosphorimaging as described in Materials and Methods. The migration positions of the helicase substrate (S) and the displaced 50mer reaction product (P) are indicated to the left. The letters above the lanes are: BC, before column sample; FT, flow through sample after loading. The numbers above the lanes indicate the fraction numbers. (B) Aliquots of the TK119/RNA–Sepharose column fractions were resolved by SDS–PAGE and polypeptide bands were detected by silver staining, as described in Materials and Methods. (C) Aliquots of the TK119/RNA–Sepharose column fractions were resolved by 10% SDS–PAGE gels and western analysis was carried out as described in Materials and Methods, using a monoclonal antibody against the human hnRNP L protein. For (B) and (C), the letters above the lanes are: BC, before column sample; FT, flow through sample after loading; M, molecular weight markers with the known kDa position indicated to the left. The numbers above the lanes indicate the fraction numbers. For (A)–(C), fractions 4–20 eluted with 75 mM KCl, and fractions 24–40 eluted with 150 mM KCl.