To quantify binding, we employed our previously described implementation of Cram's extraction method.[ 7, 18, 19] Briefly, the receptor is dissolved in chloroform then equilibrated with an aqueous solution of Et4N+X−, where X− is the anion of interest.
To determine the order of calcium-binding and apparent KD values for each of the individual calcium-binding sites, we employed a crystallographic approach.
To investigate genome-wide binding of Drosha, we employed a ChIP-on-chip approach using human 5.1.1 ENCODE array.
To characterize further the effect of the T63S mutation on ion binding to the channel, we employed a biochemical assay [15] in which the effects of potassium and barium ions on the stability of the channel tetramer was examined (Figure 6).
To verify the specific binding of SH122 to IAPs, we employed a pull-down assay with biotin-labeled SH122 (SH122BL) in human prostate cancer cells.
To confirm the importance of the octanoyl chain for membrane-binding, we employed another technique as well as another membrane-mimetic.
Given that we employed a peptide binding motif, imposing certain restrictions for the p1, p4, p6 and p9 positions, for the identification of this T cell epitope within HC gp-39 [ 7, 41], some more impact of the single site substitutions at p4, p6 and p9 was anticipated.
To test whether netrin-CD146 interaction could occur on the surface of living cells, we employed a cell surface binding assay.
Next we employed a GST in vitro binding assay to confirm the physical interaction between Zfp277 and Bmi1 or Mel18 using GST-Bmi1 and GST-Zfp277, respectively.
