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. Author manuscript; available in PMC: 2011 Aug 27.
Published in final edited form as: Chem Biol. 2010 Aug 27;17(8):795–801. doi: 10.1016/j.chembiol.2010.05.027

Figure 2. Activity-Based Profiling, Affinity Purification, and LC-MS Identification of Proteasome β Subunits in Murine Tissues Lysates.

Figure 2

(A) In-gel fluorescence detection of active proteasome β subunits in 3-week-old wild-type murine thymus homogenate after labeling with MV151, ABP 1, 2, and 4 (see also Figure S1). M indicates the molecular marker band of 25 kDa.

(B) In-gel fluorescence and silver stain detection of active proteasome β subunits in young and adult thymus after labeling with ABP 1, 2, 3, and affinity purification. Protein identification by LC-MS analysis of in-gel digested silver-stained bands (see Tables S1 and S2 for details).

(C) In-gel fluorescence detection with ABP 4 of β5t activity in wild-type and absence of activity in the (−/−) β5t knockdown thymus from 3-weeks-old mice.

(D) Activity-based proteasome profiling using ABP 4 shows β5t activity in murine thymus (young and adult) but not in heart, lung, liver, spleen, brain, testes, or kidney.

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