Methods are described for localizing proteins in unstained gels and accurately excising the regions containing them. A gel elutor, which is all glass with Lucite fittings, is also described. The elutor removes proteins from gels with a high yield and concentrates the protein in a small volume. The elutor is simple and very easy to use. A way is presented for avoiding the oxidation of methionine and cysteine during preparative electrophoresis.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1016/0003-2697(78)90811-4 | DOI Listing |
Nat Protoc
May 2020
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA, USA.
In pathology, microscopy is an important tool for the analysis of human tissues, both for the scientific study of disease states and for diagnosis. However, the microscopes commonly used in pathology are limited in resolution by diffraction. Recently, we discovered that it was possible, through a chemical process, to isotropically expand preserved cells and tissues by 4-5× in linear dimension.
View Article and Find Full Text PDFJ Chromatogr B Analyt Technol Biomed Life Sci
March 2019
Institute of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, PR China. Electronic address:
Proteomic analysis by combining PAGE separation, gel slicing and slice-by-slice in-gel digestion and LC-MS/MS has been frequently reported in recent years. Since the MS analysis would provide identities and quantities of the proteins along the whole lane, gel visualization by dye staining could be skipped to save time and labor. In this work, we examined the effects of CBB R-250 staining on the performance of the method and the results showed actually better results were obtained with CBB staining than without, both in nondenaturing PAGE and SDS-PAGE.
View Article and Find Full Text PDFJ Oncol Res Ther
February 2018
Department of Comparative Pathobiology, Purdue University Center for Cancer Research, Purdue University, Indiana, USA.
Laser Capture Micro-dissection (LCM) is a technique that is used to isolate specific tumor cells from a heterogeneous tumor tissue sample.To aid in identifying and dissecting pure tumor cells from other parts of the tissues such as the stroma, tissues are stained with Haematoxylin and Eosin. The cells are then used for protein, RNA or DNA extraction.
View Article and Find Full Text PDFIsolated thylakoid membranes were disrupted by treatment with nonionic detergents digitonin or dodecyl maltoside. Solubilized polypeptide complexes were separated by native gel charge shift electrophoresis. The position of ATP-synthase complex and its isolated catalytic part (CF1) within gel was determined using the color reaction for ATPase activity.
View Article and Find Full Text PDFMethods Mol Biol
March 2016
Departament de Bioquímica i Biologia Molecular, Facultat de Biociències, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, 08193, Spain.
This chapter describes very simple fluorescent methods developed in our laboratory allowing the rapid monitoring of total protein patterns on both sodium dodecyl sulfate (SDS) polyacrylamide gels and western blots. The noncovalent dye Nile red (9-diethylamino-5H-benzo[α]phenoxazine-5-one) is used for the sensitive staining of proteins in SDS gels. This method is compatible with the electroblotting of protein bands and with the staining of the resulting blot with the covalent dye MDPF (2-methoxy-2,4-diphenyl-3(2H)-furanone).
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!