Journal:

Article Title: Nitrated alpha-synuclein and microglial neuroregulatory activities

doi: 10.1007/s11481-008-9100-z

Figure Lengend Snippet: SELDI-TOF, 1D SDS PAGE, and Western blot analyses of supernatant fluids obtained from N-α-syn-activated microglial. Representative SELDI-TOF spectral analysis (region 10–20 kDa) of untreated-control (top panel) and N-α-syn-stimulated microglia (bottom panel) at 16 h post-stimulation, shown in A. Marked by an asterisk are upregulated and uniquely expressed peaks corresponding in molecular weight within 1% of mass tolerance to proteins identified by LC-MS/MS. These include calcyclin (10,051 Da), thioredoxin (11,544 Da), calvasculin (11,721 Da), calmodulin (16,706 Da), and TNF-α (17,907 Da). Bands were excised from 1D SDS PAGE gel, digested by trypsin, and sequenced by LC-MS/MS. Lanes are supernatant fluids obtained from control (unstimulated) microglial = [Lanes 1–3] and supernatants from N-α-syn-activated microglia [Lanes 4–6] collected 8 h, 16 h, and 24 h post-stimulation, respectively. (B). Representative Western blots of supernatant fluids from control and N-α-syn-stimulated microglia 16 h post-stimulation for proteins identified by LC-MS/MS (C).

Article Snippet: Protein profiling of culture supernatants was performed using SELDI-TOF ProteinChip ® assays ( Enose et al., 2005 ; Kadiu et al., 2007 ) (Ciphergen Biosystems, Fremont, CA).

Techniques: SDS Page, Western Blot, Molecular Weight, Liquid Chromatography with Mass Spectroscopy