The very sensitive and reliable silver staining method to visualize proteins in polyacrylamide gels described by Wray et al. (Anal. Biochem. (1981) 118, 197-203) fails when the protein sample contains nucleic acids and/or metals. By washing the polyacrylamide gels in acetic acid and repeatedly in methanol immediately following electrophoresis and then using the procedure of Wray et al., many gels otherwise unstainable may be stained with a high degree of reliability. This method allows visualization of a minute amount of proteins in samples containing high amounts of DNA and metals.
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http://dx.doi.org/10.1016/0003-2697(86)90349-0 | DOI Listing |
J Proteome Res
January 2025
Advanced Research Support Center, Ehime University, Ehime 791-0295, Japan.
Precise prefractionation of proteome samples is a potent method for realizing in-depth analysis in top-down proteomics. PEPPI-MS (Passively Eluting Proteins from Polyacrylamide gels as Intact species for MS), a gel-based sample fractionation method, enables high-resolution proteome fractionation based on molecular weight by highly efficient extraction of proteins from polyacrylamide gels after SDS-PAGE separation. Thereafter it is essential to effectively remove contaminants such as CBB and SDS from the PEPPI fraction prior to mass spectrometry.
View Article and Find Full Text PDFNat Protoc
January 2025
Advanced Research Support Center, Ehime University, Ehime, Japan.
Top-down analysis of intact proteins and middle-down analysis of proteins subjected to limited digestion require efficient detection of traces of proteoforms in samples, necessitating the reduction of sample complexity by thorough pre-fractionation of the proteome components in the sample. SDS-PAGE is a simple and inexpensive high-resolution protein-separation technique widely used in biochemical and molecular biology experiments. Although its effectiveness for sample preparation in bottom-up proteomics has been proven, establishing a method for highly efficient recovery of intact proteins from the gel matrix has long been a challenge for its implementation in top-down and middle-down proteomics.
View Article and Find Full Text PDFEur J Obstet Gynecol Reprod Biol
February 2025
Department of Obstetrics and Gynecology, Aalborg University Hospital (AAUH), Denmark.
Introduction And Hypothesis: Polyacrylamide hydrogel (PAHG) is a bulking agent used in treatment of stress urinary incontinence (SUI) in women. Some women experience an improvement or cure in stress urinary incontinence (SUI) symptoms lasting only a few weeks after the injection of the bulking agent. The aim of this study was to use three-dimensional ultrasound (3DUS) to describe the changes in volume, number of deposits and echogenicity of the bulking agent 3 months after PAHG injection.
View Article and Find Full Text PDFSoft Matter
December 2024
Department of Macromolecular Science and Engineering, Case Western Reserve University, Cleveland, OH, USA.
Collagen type II fibrils provide structural integrity to the articular cartilage extracellular matrix. However, the conditions that control the fibril radial size scale, distribution, and formation inside of dense networks are not well understood. We have investigated how surrounding elastic networks affect fibril formation by observing the structure and dynamics of collagen type II in model polyacrylamide gels of varying moduli.
View Article and Find Full Text PDFReprod Domest Anim
November 2024
Department of Veterinary Clinical Complex, West Bengal University of Animal & Fishery Sciences, Kolkata, India.
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