gel electrophoresis 119 2de  (Bio-Rad)

Journal: Technology in cancer research & treatment

Article Title: Identification of Differentiation-Related Proteins in Gastric Adenocarcinoma Tissues by Proteomics.

doi: 10.1177/1533034615595792

Figure Lengend Snippet: Figure 3. Enlarged 2-dimensional gel electrophoresis (2DE) images of altered proteins. Proteins upregulated in PDGA compared with well- differentiated gastric adenocarcinoma (WDGA) included calreticulin precursor (spot 1), tapasinERP57 heterodimer (spot 2), pyruvate kinase type M2 (PKM2; spot 3), class Pi glutathione S-transferase (GST-Pi; spot 4), and enolase 1 (ENO1; spots 5 and 6). Proteins were downregulated in PDGA included myosin-11 isoform SM2A (spot 7) and actin alpha cardiac (spot 8). Each spot number is the same as those in Figure 2. PDGA indicates poorly differentiated gastric adenocarcinoma.

Article Snippet: The power supply, the 2-dimensional (2D) gel electrophoresis (2DE) system (PROTEAN IEF cell, PROTEAN II Xi cell), the Trans-Blot Cell, immobilized pH gradient (IPG) strip (pH 3-10, nonlinear, 17 cm), Bio-Lyte (pH 3-10), urea, CHAPS, sodium dodecyl sulfate (SDS), acrylamide, N,N0-methylenebisacrylamide, glycine, and Tris-base were purchased from Bio-Rad (Richmond, California).

Techniques: Nucleic Acid Electrophoresis

Journal: Technology in cancer research & treatment

Article Title: Identification of Differentiation-Related Proteins in Gastric Adenocarcinoma Tissues by Proteomics.

doi: 10.1177/1533034615595792

Figure Lengend Snippet: Figure 2. Representation of 2-dimensional gel electrophoresis (2DE) analysis of proteome profiles of gastric cancer. Total protein (800 mg) extracted from (A) PDGA tissues and (B) WDGA tissues was separated on nonlinear strips with a pH range of 3 to 10, followed by protein separation by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). In all, 6 spots (1, 2, 3, 4, 5, and 6) were upregulated and 2 spots (7 and 8) were downregulated in PDGA tissues compared with WDGA. PDGA indicates poorly differentiated gastric adenocar- cinoma; WDGA, well-differentiated gastric adenocarcinoma.

Article Snippet: The power supply, the 2-dimensional (2D) gel electrophoresis (2DE) system (PROTEAN IEF cell, PROTEAN II Xi cell), the Trans-Blot Cell, immobilized pH gradient (IPG) strip (pH 3-10, nonlinear, 17 cm), Bio-Lyte (pH 3-10), urea, CHAPS, sodium dodecyl sulfate (SDS), acrylamide, N,N0-methylenebisacrylamide, glycine, and Tris-base were purchased from Bio-Rad (Richmond, California).

Techniques: Nucleic Acid Electrophoresis, Polyacrylamide Gel Electrophoresis, SDS Page

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Redox Regulatory Mechanism of Transnitrosylation by Thioredoxin *

doi: 10.1074/mcp.M110.000034

Figure Lengend Snippet: 2DE analysis of biotinylated proteins from vector (V ec )- ( A ) and Trx1 C32S/C35S ( B )-transfected HeLa cells is shown. Nitrosylated proteins were biotinylated, enriched and eluted from streptavidin beads, desalted with ice-cold acetone, dissolved in a 2DE rehydration buffer, and analyzed by 2DE. Gels were stained with SYPRO Ruby dyes, and the protein spots whose densities were increased in Trx1 C32S/C35S -transfected cells were excised for trypsin digestion and protein identification by tandem MS ( supplemental Table SI ). C , proteins recovered from streptavidin beads were analyzed by Western blotting with individual antibodies specific for the detection of nitrosylated Trx1, Prx1, HSP90β, actin, Casp3, Txnip, and TrxR. Values are the mean ± S.E. for experiments performed in triplicate (*, p = 0.0006; †, p = 0.001; ‡, p = 0.0001; §, p = 0.0007; , p = 0.001; #, p = 0.0009; **, p = 0.002; Student's t test). Non-streptavidin-enriched proteins were analyzed in parallel to detect proteins regardless of their nitrosylation status.

Article Snippet: Two-dimensional gel electrophoresis (2DE) materials and SYPRO Ruby stain were purchased from Bio-Rad.

Techniques: Plasmid Preparation, Transfection, Staining, Western Blot