Figure 1.

Identification of albumin as a binding partner for the NW peptide. (A) Experimental workflow of the chemical cross-linking. In this protocol, monocytes were first incubated with the biotinylated NW peptide and then the chemical cross-linker DSP was added to stabilize the interaction between the NW peptide and its binding receptor(s). After cell lysis, membrane proteins were prepared and streptavidin magnetic beads were applied to pull down the peptide–protein complexes prior to SDS-PAGE analysis. (B) Analysis of the eluted proteins by SDS-PAGE. After electrophoresis, the gel was silver stained. As a control, monocytes were incubated with DSP only. A single sharp band was identified in the sample incubated with both the biotinylated NW peptide and the DSP cross-linker. (C) An additional experiment. Experimental conditions are as in (B). The data shown in (B,C) are representative of six independent experiments. DSP, Dithiobis(succinimidyl propionate); NW, refers to the peptide’s name; SDS-PAGE, Sodium dodecyl-sulfate polyacrylamide gel electrophoresis.