Journal: PLoS ONE

Article Title: Increased Phosphorylation of Vimentin in Noninfiltrative Meningiomas

doi: 10.1371/journal.pone.0009238

Figure Lengend Snippet: Two-dimensional hierarchical clustering of 64 tumors (32 infiltrative, two invasive, and 30 noninvasive tumors) was performed with 69 molecular mass peaks after SELDI-TOF mass spectra processing with the Biomarker Wizard software (Ciphergen). Candidate markers and patient samples were clustered using complete linkage clustering methods from Eisen's cluster software. Clustered trees are displayed using Eisen's Treeview software. Red squares denote high marker concentration in comparison to average; green squares denote low concentration in comparison to average. The numbers of infiltrative or invasive samples and of noninfiltrative or noninvasive samples in the two clusters are indicated.

Article Snippet: After 30 min incubation on ice and centrifugation (10,000 g for 10 min at 4°C), supernatant was diluted in a binding buffer (100 mM Tris and 0.1% TritonX100 at pH 8.0) to a final protein concentration of 0.1 μg/μL, and 100 μL of this suspension was applied to Q10 anion-exchange active binding surfaces of SELDI ProteinChip Arrays (Bio-Rad, Marnes-La-Coquette, France).

Techniques: Biomarker Discovery, Software, Marker, Concentration Assay, Comparison

Journal: PLoS ONE

Article Title: Increased Phosphorylation of Vimentin in Noninfiltrative Meningiomas

doi: 10.1371/journal.pone.0009238

Figure Lengend Snippet: Tumor extracts were analyzed using SELDI-TOF on Q10 anion-exchange ProteinChip Arrays, as shown in . Signal intensity at the 53-kDa mass level were measured and calculated parameters plotted for (A) infiltrative/invasive tumor samples, (B) noninfiltrative tumor samples. Medians, 25 th and 75 th percentiles are marked with line segments across the boxes and the lowest and highest signal values with bars.

Article Snippet: After 30 min incubation on ice and centrifugation (10,000 g for 10 min at 4°C), supernatant was diluted in a binding buffer (100 mM Tris and 0.1% TritonX100 at pH 8.0) to a final protein concentration of 0.1 μg/μL, and 100 μL of this suspension was applied to Q10 anion-exchange active binding surfaces of SELDI ProteinChip Arrays (Bio-Rad, Marnes-La-Coquette, France).

Techniques:

Journal: PLoS ONE

Article Title: Increased Phosphorylation of Vimentin in Noninfiltrative Meningiomas

doi: 10.1371/journal.pone.0009238

Figure Lengend Snippet: After purification by chromatography and electrophoresis, the 53-kDa marker was cleaved by proteases. (A) peptide fingerprint after GluC endoproteinase digestion obtained by SELDI-TOF analysis on NP20 ProteinChip Arrays. Peptides with measured molecular masses matching those of the computed GluC endoproteinase proteolytic peptides of vimentin are indicated with an asterisk. (B) GluC endoproteinase peptides of vimentin identified by SELDI-TOF are listed according to their masses. (C) Mapping of the peptides identified by either peptide mass fingerprinting or nanoLC-MS/MS to the vimentin sequence. Sequences highlighted in grey correspond to GluC endoproteinase peptides identified in B. Underlined sequences correspond to 60 trypsin peptides identified by nanoLC-MS/MS. Phosphorylated peptides (37-50 amino acid and 70-78 amino acid peptides) are underlined with dots.

Article Snippet: After 30 min incubation on ice and centrifugation (10,000 g for 10 min at 4°C), supernatant was diluted in a binding buffer (100 mM Tris and 0.1% TritonX100 at pH 8.0) to a final protein concentration of 0.1 μg/μL, and 100 μL of this suspension was applied to Q10 anion-exchange active binding surfaces of SELDI ProteinChip Arrays (Bio-Rad, Marnes-La-Coquette, France).

Techniques: Purification, Chromatography, Electrophoresis, Marker, Peptide Mass Fingerprinting, Tandem Mass Spectroscopy, Sequencing

Journal: PLoS ONE

Article Title: Increased Phosphorylation of Vimentin in Noninfiltrative Meningiomas

doi: 10.1371/journal.pone.0009238

Figure Lengend Snippet: Purified 53-kDa marker from noninfiltrative tissue extracts was treated by alkaline phosphatase and samples were analyzed using SELDI-TOF MS. Analyses were performed on Q10 anion-exchange ProteinChip Arrays (A and B) or hydrophilic NP20 ProteinChip Arrays (C and D). (A and C) Controls with untreated purified 53-kDa marker. (B and D) phosphatase-treated marker.

Article Snippet: After 30 min incubation on ice and centrifugation (10,000 g for 10 min at 4°C), supernatant was diluted in a binding buffer (100 mM Tris and 0.1% TritonX100 at pH 8.0) to a final protein concentration of 0.1 μg/μL, and 100 μL of this suspension was applied to Q10 anion-exchange active binding surfaces of SELDI ProteinChip Arrays (Bio-Rad, Marnes-La-Coquette, France).

Techniques: Purification, Marker

Journal: International Journal of Molecular Sciences

Article Title: Mass Spectrometry-Based Proteomics for Pre-Eclampsia and Preterm Birth

doi: 10.3390/ijms160510952

Figure Lengend Snippet: The Applications of MALDI-MS and SELDI-MS for Preeclampsia and Preterm Birth.

Article Snippet: SELDI-MS (Q10 array) , Amniotic fluid (AF) , PE: 18 CHTN: 7 CRL: 16 , Before sample loading, each ProteinChip spot was incubated twice with 5 μL binding buffer in a humidity chamber at room temperature for 5 min. After equilibration, 5 μL of sample, diluted 1:3 in binding buffer, was added to each spot and incubated in a humidity chamber with shaking for 40 min. Each spot was washed with 5 μL binding buffer for 2 min, followed by washing with 5 μL of triple distilled water, and air-dried. , 2 peaks located at 17,399.1 and 28,023.3 Da were significantly different. The former peak distinguished women with PE from control, and the latter peak distinguished women with PE and CHTN from controls. The peaks were assigned to hypothetical protein SBBI42 and proapolipoprotein A-I. The results were cross-validated with Western blot. , It was suggested that the increase in levels of proapolipoprotein A-I in the AF of women with PE may represent a compensatory mechanism to maintain levels of apolipoprotein A-I and thereby pulmonary surfactant and lung compliance and development. , [ ] .

Techniques: Sample Prep, Extraction, Magnetic Beads, Incubation, Binding Assay, Control, Modification, Clinical Proteomics, Synthesized, Enzyme-linked Immunosorbent Assay, Spectrophotometry, Western Blot, Expressing, Activity Assay, Concentration Assay, Infection, Derivative Assay, Generated, Sterility