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Figure 5 | Proteome Science

Figure 5

From: SELDI-TOF-MS ProteinChip array profiling of T-cell clones propagated in long-term culture identifies human profilin-1 as a potential bio-marker of immunosenescence

Figure 5

Western blot analysis of profilin-1 expression in T-cell clones obtained from octogenarian donors. For Western blot analysis, one million cells were pelleted and lysed in sample buffer (50 mM Tris, pH 8.0, 120 mM NaCl, 0.5% NP40, 10 μg/mL PMSF (phenylmethylsulfonylfluoride), and 1× protease inhibitor cocktail. A total of 30 μg protein was suspended in laemmli buffer (50 mM tris, pH 6.8, 1% β-mercaptoethanol, 2% sodium dodecyl sulphate, 0.1% bromophenol blue, and 10% glycerol). Samples were boiled and subject to Western blot analysis on NuPAGE™ Bis-Tris gels with MES running buffer. Human profilin-1, cofilin and β-actin proteins were detected by antibodies as detailed in Materials and Methods and visualised by enhanced chemi-luminescence.

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Proteome Science

ISSN: 1477-5956