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. Author manuscript; available in PMC: 2014 May 28.
Published in final edited form as: Biochemistry. 2013 May 16;52(21):10.1021/bi301662w. doi: 10.1021/bi301662w

Figure 7.

Figure 7

(A) Chemical structure of amine- reactive, homobifunctional, and isotope-labeled (do and d4) cross-linking reagent BS2G - Bis (sulfosuccinimidyl) glutarate. The corresponding mass shifts associated with d0 and d4 cross-linked peptides are shown. (B) SDS-PAGE of αB-crystallin. Biotinylated αA66-80 peptide cross-linking reaction mixture with BS2G d0/d4 and western blot of similarly run gel. The reaction was carried out as described under methods with 20-fold molar excess for 45 min. SDS-PAGE and Western blot: Lane 1- Marker, Lane 2- αBWT control with cross-linker (without peptide), Lane 3- αBWT + Biotin-αA66-80 without cross-linker, Lane 4- αBWT + Biotin-αA66-80 with cross-linker. Arrows in the SDS-PAGE gel point to cross-linked complexes containing biotin-labeled peptide confirmed by western blot of the gel.