A convenient method was established for the M(r) estimation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of large polypeptides in the range from 250,000 to 600,000. Such polypeptides had previously been shown to migrate improperly in various gel electrophoresis systems. In a conventional Laemmli system with 3% polyacrylamide, approximately linear log M(r) vs Rf plots could be obtained by varying the gel thickness which ranged from 0.4 to 1.5 mm. In the additional presence of an electrolyte, such as NaCl, running times were shorter and for evaluation linear plots could be constructed from runs in very thin gels with less than 0.5 mm thickness. Handling of gels was greatly facilitated by fixing them to one of the gel chamber glass plates via bind-silane treatment prior to casting.
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http://dx.doi.org/10.1006/abio.1993.1527 | DOI Listing |
Nucleic Acids Res
January 2025
Department of Pharmaceutical Sciences, University of California, Irvine, CA 92697-3958, United States.
Growth in the development of engineered polymerases for synthetic biology has led to renewed interest in assays that can measure the fidelity of polymerases that are capable of synthesizing artificial genetic polymers (XNAs). Conventional approaches require purifying the XNA intermediate of a replication cycle (DNA → XNA → DNA) by denaturing polyacrylamide gel electrophoresis, which is a slow, costly, and inefficient process that requires a large-scale transcription reaction and careful extraction of the XNA strand from the gel slice. In an effort to streamline the assay, we developed a purification-free approach in which the XNA transcription and reverse transcription steps occur inside the matrix of a hydrogel-coated magnetic particle.
View Article and Find Full Text PDFJ Chromatogr A
January 2025
Waters Corporation, Instrument/Core Research/Fundamental, Milford, MA, 01757, USA. Electronic address:
Significant progress has been made in the last two decades in producing small (<2μm), high-purity, and low-adsorption particles, columns and system hardware, for ultra-high pressure liquid chromatography (UHPLC). Simultaneously, the recent rapid expansion of cell and gene therapies for treating diseases necessitates novel analytical technologies for analyzing large (>2 kbp) plasmid double-stranded (ds) DNA (which encodes for the in vitro transcription (IVT) of single-stranded (ss) mRNA therapeutics) and dsRNAs (related to IVT production impurities) biopolymers. In this context, slalom chromatography (SC), a retention mode co-discovered in 1988, is being revitalized using the most advanced column technologies for improved determination of the critical quality attributes (CQAs) of such new therapeutics.
View Article and Find Full Text PDFUrol Res Pract
January 2025
Department of Pediatric Surgery, Zonguldak Bulent Ecevit University, Faculty of Medicine, Zonguldak, Türkiye.
Objective: Bladder tissue models have been developed using smooth muscle cells (SMCs) on various scaffolds to mimic bladder morphology and physiology. This study investigates the effects of co-culturing fetal and adult SMCs on growth properties and protein profiles to understand cellular interactions and population kinetics.
Methods: Bladder tissue samples from 10 adult and 10 fetal New Zealand rabbits were divided into 5 groups: adult SMCs (A), fetal SMCs (F), 50%A+50%F (A+F), 75%A+25%F (3A+F), and 25%A+75%F (A+3F).
J Dent Sci
January 2025
School of Dentistry and Institute of Oral Medicine, National Cheng Kung University, Tainan, Taiwan.
Background/purpose: The efficacy of riboflavin-ultraviolet-A (RF-UVA) treatment in crosslinking collagen and improving dentin bonding has been proven. However, biodegradation of the hybrid layer may compromise the bonding. The purpose of this study was to evaluate different RF-UVA treatments regarding their ability to preserve dentin bonding from enzymatic digestion.
View Article and Find Full Text PDFPeerJ
January 2025
Department of Biology, School of Sciences and Humanities, Nazarbayev University, Astana, Kazakhstan.
Background: Chitosan nanoparticles (CsNPs) are an effective and inexpensive approach for DNA delivery into live cells. However, most CsNP synthesis protocols are not optimized to allow long-term storage of CsNPs without loss of function. Here, we describe a protocol for CsNP synthesis, lyophilization, and sonication, to store CsNPs and maintain transfection efficiency.
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