Skip to main content
. Author manuscript; available in PMC: 2013 May 4.
Published in final edited form as: J Proteome Res. 2012 Apr 4;11(5):2721–2738. doi: 10.1021/pr2010814

Figure 2. Use of SILAC internal standards to quantify the detergent insoluble brain proteome.

Figure 2

(A) SDS-PAGE analysis of Brain/SILAC mixtures. Molecular weight marker (M; lane 1), whole cell lysate of SILAC labeled HEK293 cells (lane 2), FTLD brain insoluble-proteome alone (lane 3), FTLD/SILAC mixture (lane 4), control brain insoluble-proteome alone (lane 6), Control/SILAC mixture (lane 7). 20 μg of protein loaded in each lane. The brain/SILAC samples (lanes 4 and 7) were, excised into gel slices based on MW (regions A-E), digested with trypsin, and analyzed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) on a hybrid linear ion-trap/Orbitrap mass spectrometer. (B) The SILAC labeled peptides from the HEK293 cells are chemically identical to their native/unlabeled counterparts and serve as internal standards for the measurement of protein abundance across control and FTLD samples. (C) Representative base peak elution profiles (MW region A) of control:HEK293 and FTLD:HEK293 peptide mixtures, respectively.