Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Serum Amyloid A Facilitates the Binding of High-Density Lipoprotein From Mice Injected With Lipopolysaccharide to Vascular Proteoglycans

doi: 10.1161/atvbaha.111.226159

Figure Lengend Snippet: Figure 2. MALDI-TOF analysis. HDL (d1.063 to 1.210 g/mL) was isolated by ultracentrifugation from plasma of control and LPS-injected mice. HDL (20 g of protein) was separated by SDS-PAGE (10% to 20% gradient gel), and the gel was stained with Coomassie Brilliant Blue. Each gel band corresponding to the apparent molecular weight of SAA1.1/2.1, SAA4, apoA-I, and apoE was cut out and digested with trypsin, and the pep- tide digest was extracted for tandem mass spectrometric analy- sis by MALDI-TOF. The arrows indicate bands that were identi- fied by MALDI-TOF and database searching that contained peptides unique to SAA1.1/2.1, SAA4, apoA-I, and apoE (A). Albumin-depleted plasma samples (20 L) from control (B-upper panel) and LPS-injected (B-lower panel) mice were separated by 2-dimensional electrophoresis (first dimension: IEF pH 3 to 10; second dimension: 10% SDS-PAGE), and the gel was stained with a silver stain. Selected spots from 2-dimensional gels were identified by in-gel tryptic digest and MALDI-TOF analysis. The small arrows indicate bands that were identified by tandem mass spectrometry MALDI-TOF and database searching that contained peptides unique to SAA1.1 (small arrow B), SAA2.1 (small arrow A), and apoA-I (B). The spot designated SAA2.1 was identified as such with a MASCOT MOWSE score of 374 (CI 100%) (C), and the adjacent spot was identified as SAA1.1 with a MOWSE score of 403 (CI 100%) (D). *Peaks corresponding to SAA peptides.

Article Snippet: Plasma samples (20 μl), from which albumin was removed using Seppro Human Albumin IgY (GenWay Biotech, Inc. San Diego, CA), were separated by two dimensional electrophoresis (first dimension: IEF pH 3-10, second dimension: 10% SDS- PAGE, Bio-Rad Laboratories, Hercules, CA) as per the manufacturer’s instructions.

Techniques: Isolation, Clinical Proteomics, Control, Injection, SDS Page, Staining, Molecular Weight, Electrophoresis, Silver Staining, Mass Spectrometry

Journal: Proteome Science

Article Title: Effect of long-term exposure of SH-SY5Y cells to morphine: a whole cell proteomic analysis

doi: 10.1186/1477-5956-4-23

Figure Lengend Snippet: 2-DE pattern of untreated (A) and 6 h morphine-treated (B) SH-SY5Y cells . Sample were resolved by 2-DE on non-linear pH 3–10 IPG strips followed by separation on a 12% SDS-PAGE gel in the second dimension. Proteins were visualized by colloidal coomassie staining. The box in 1B delineates the close-up presented on figure 2.

Article Snippet: 1.2 μl of IPG buffer pH 3–10 NL (Amersham Biosciences) and 2 μl of 1% (w:v) orange G were added to 250 μl of sample before application to the strip.

Techniques: SDS Page, Staining