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. 2015 May 13;6:315. doi: 10.3389/fpls.2015.00315

Figure 2.

Figure 2

SDS-PAGE analysis of proteins extracted from the late growth stage Arabidopsis stems for the in-solution and in-gel digestion approaches in identifying stem proteins. (A) SDS-PAGE analysis with a range of protein loading amount, Lanes 1–3, the three biological replicate samples used for in-solution digestion analysis; 10 μg proteins per well. Lanes 4 and 5, the two biological replicate samples used for in-gel digestion analysis; 30 μg proteins per well. M: protein marker (#P7702, New England Biolabs, USA); NuPage 12% Bis-Tris gel was used for better separation of small- to medium-size proteins (15–75 kDa). (B) SDS-PAGE separation of proteins used for in-gel digestion analysis, with 30 μg per well. M: protein marker (#P7703, New England Biolabs, USA). NuPage 4–12% Bis-Tris gel was used for better separation of medium- to large-size proteins (30–250 kDa), which resulted in 20 distinguishable strong bands (S0–S19, indicated by red boxes) and 25 weak bands (W1–W25; covering the remaining gel areas after strong bands were removed), which were used for digestion and subsequent LC-MS analysis. Note that the band S0 is the location of protein-loading well.