Search results for "Trypsinization"
showing 5 items of 5 documents
Cell Harvesting Methods Affect Cellular Integrity of Adherent Cells During Apoptosis Detection.
2018
Background/aim Annexin V and propidium iodide (PI) dual staining is commonly applied in bioscience as a method to detect apoptosis. However, excessive handling of adherent cells may interfere with the integrity of plasma membrane and hence impede the accuracy of this method. Here, we exploited PI uptake as an indicator of cell integrity and investigated how cell harvesting methods and solutions involved in common apoptosis detection techniques affected measurement results. Materials and methods Different cell harvesting techniques, staining with PI and flow cytometry were performed. Results Non-fixed scrapped cells revealed significantly higher fractions of PI-positive staining compared to …
Scanning electron microscopy of heterochromatin in chromosome spreads of male germ cells in Schistocerca gregaria (Acrididae, Orthoptera) after tryps…
1996
Chromosome spreads, prepared from testes of the desert locust Schistocerca gregaria, were analyzed using scanning electron microscopy (SEM) after varying periods of preincubation in trypsin. The emphasis of the study was on the appearance of heterochromatin. A trypsin pretreatment of 5 sec resulted in a smooth surface on the chromatin throughout and the heterochromatin was highly electron-emissive. The facultatively heterochromatic X chromosome was clearly visible in interphase spermatogonia and in pachytene and late prophase I spermatocytes. Chromomeres of autosomal bivalents could be recognized in pachytene cells. Centromeric heterochromatin segments were very prominent in autosomes of la…
Western blotting as a method for studying cell-biomaterial interactions: The role of protein collection
2000
Research of cell-biomaterial interactions is building on knowledge and methods available in cell and molecular biology. Western blotting is one of the options to characterize protein expression in cell populations. Method transfer to biomaterial model systems is not trivial because of the structure that exists in many biomaterials, preventing the collection of cell lysate by mechanical means. In this technical report, we describe the influence of different protein collection methods in a model system for cell-biomaterial interactions, consisting of endothelial cells exposed to different stimuli. In particular, the influence of trypsinization before lysis, and handling complexity were determ…
The yeast histone acetyltransferase A2 complex, but not free Gcn5p, binds stably to nucleosomal arrays.
2000
We have investigated the structural basis for the differential catalytic function of the yeast Gcn5p-containing histone acetyltransferase (HAT) A2 complex and free recombinant yeast Gcn5p (rGcn5p). HAT A2 is shown to be a unique complex that contains Gcn5p, Ada2p, and Ada3p, but not proteins specific to other related HAT A complexes, e.g. ADA, SAGA. Nevertheless, HAT A2 produces the same unique polyacetylation pattern of nucleosomal substrates reported previously for ADA and SAGA, demonstrating that proteins specific to the ADA and SAGA complexes do not influence the enzymatic activity of Gcn5p within the HAT A2 complex. To investigate the role of substrate interactions in the differential …
The lipoprotein lipase activity in cultured beating heart cells of the post-natal rat.
1974
Summary The lipoprotein lipase (LPL) activity was studied in cultured beating heart cells of the post natal rat as a function of the culture age, from the freshly trypsinized cell suspension up to the 14th day of culture. The LPL activity remains at a practically stationary level during the first four days of culture, and then increases rapidly to reach a plateau, at a level 3–4 times higher at the 21th day of culturing. The significance of this enzymatic activity progression comparing to 3H thymidine incorporation from the point of view of cell differentiation is discussed. Our results suggest that after the 12th day of culture, cells contain their optimal enzymatic apparatus for lipid met…