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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: J Clin Periodontol. 2014 Jun 6;41(8):733–747. doi: 10.1111/jcpe.12262

Figure 2. SDS-PAGE electroelution of GCF proteins from periopaper strips from healthy and periodontal subjects for stable-isotope labeling and quantitative proteome analysis by MS.

Figure 2

(A) Short time SDS-PAGE run: a single periopaper containing ~0.2–0.7 μl GCF were placed in each well containing sample buffer. The SDS-PAGE was run until the dyefront was approximately half-way the gel length. This was sufficient to electroelute proteins from the periopaper into the gel visualized by Coomassie blue staining. Molecular weight standards can be seen on the left side of the gel. The electroeluted GCF proteins are not visible at this point since the gel has not been yet stained with coomassie blue. (B) Long time SDS-PAGE run: 4 periopaper strips derived from healthy individuals and 4 periopapers derived from periodontal patients, respectively, were run in a single SDS-PAGE separated by an empty lane. After comassie staining the gel was sectioned into different molecular weight regions as indicated. Lane 1, standard molecular weight proteins; Lanes 2–5, 4 individual periopapers from healthy and lanes 7–10 individual periopapers from periodontal patients run separately. On the right hand side under “CUT” are the sections of different molecular weight regions excised across and processed for MS analysis, cut 1: Mr range ~100 kDa and above, cut 2: Mr range ~40–80 kDa, cut 3: Mr range ~25–38 kDa, cut 4 ~11–24 kDa and cut 5: Mr range ~2–10 kDa.