Showing papers in "Methods in 1999"

TL;DR: The basic chromatin immunoprecipitation technique is remarkably versatile and has now been used in a wide range of cell types, including budding yeast, fly, and human cells, and it seems likely that many more studies, centered around chromatin structure and protein-DNA interactions in its native setting, will benefit from this technique.

TL;DR: Isothermal titration calorimetry (ITC) is the most quantitative means available for measuring the thermodynamic properties of a protein-protein interaction and is used to study the interaction between cytochrome c and two monoclonal antibodies.

TL;DR: The principles of deconVolution microscopy are discussed, different computational approaches for deconvolution are described, and interpretation of deconvolved images with a particular emphasis on what artifacts may arise are discussed.

TL;DR: Calcium imaging of neocortical brain slices is used to develop a method to reconstruct the action potentials occurring in a population of neurons, demonstrating that each action potential produces a stereotyped calcium transient in the somata of pyramidal neurons.

TL;DR: It is concluded that at least two processes generate intrinsic optical signals in hippocampal slices, one of which causes light scattering to change inversely with cell volume and is related to dilution of the cytoplasm, while the other, opposite in sign, may be due to mitochondrial swelling.

TL;DR: Dynamic measurements of neuronal morphology of neurons expressing green fluorescent protein (GFP) and GFP fusion proteins as well as functional imaging of calcium dynamics in individual dendritic spines are presented.

TL;DR: Proper procedures are provided for formulating MPL in an aqueous vehicle or an oil-in-water emulsion, which can be beneficial for most vaccine approaches being investigated today.

TL;DR: A number of techniques commonly used in the preparation and characterization of caspases are described, to facilitate the search for new components involved in cell death and to aid researchers in understanding the interactions between currently known cell death proteins.

TL;DR: Three methods for assaying polyadenylation of a specific mRNA in the context of total cellular RNA are presented, including oligo(dT)/RNase H-Northern analysis, which is the classic labor-intensive assay for polyadenolation and is included for historical reference and as a potential experimental control for the poly(A) test (PAT) assays described.

TL;DR: The use of liposomes as carriers of peptide, protein, and DNA vaccines requires simple, easy-to-scale-up technology capable of high-yield vaccine entrapment, and work from this laboratory has led to the development of techniques that can generate liposome of various sizes, containing soluble antigens such as proteins and particulate antIGens.

TL;DR: This work discusses different analysis methods used in fluctuation spectroscopy and evaluates their use for studying protein-protein interactions, and highlights some of the technical aspects.

TL;DR: Imaging IOSs in the submerged brain slice preparation provides insight into brain activity if it involves significant water movement between intracellular and extracellular compartments, including responses to osmotic imbalance, excitotoxic glutamate agonists, and oxygen/glucose deprivation, the latter leading to spreading depression.

TL;DR: Squalene or squalane emulsions without copolymers or MDP have very little toxicity and elicit potent antibody responses to several antigens in nonhuman primates and could be used to improve a wide range of vaccines.

TL;DR: Application of the tetracycline regulatory system to inducible control of a glucocorticoid receptor (GR)/GFP chimera can be very effectively controlled in this system, providing an ideal environment in which to study subcellular trafficking of the receptor and interactions with a variety of intracellular targets.

TL;DR: The development of design-based stereology provides a way to directly count or measure cells in three dimensions, avoiding errors (biases) and the need for assumptions regarding cell size, shape, and orientation to be validated, described as unbiased stereology.

TL;DR: Using multiple-label immunofluorescence confocal microscopy to characterize the cellular localization of antigens, the TSA method can be critical for double labeling with unconjugated primary antibodies raised in the same host species.

TL;DR: The theoretical and practical aspects of fluorescence polarization and its application to the study of protein-protein interactions are highlighted.

TL;DR: Iscoms prominently enhance the antigen targeting, uptake, and activity of antigen presenting cells including dendritic and B cells and macrophages resulting in the production of proinflammatory cytokines, above all interleukin (IL)-1, IL-6, and IL-12.

TL;DR: The stability of theCross-linked nucleosome in the presence of altered pH or salt/urea concentrations is described in order to indicate that there are limitations to procedures that can be used for the subsequent characterization of the cross-linked complex.

TL;DR: The reaction mechanism attempts to rationalize the relative enzymatic activities of GC, heme-deficient GC, GC-CO, and GC-NO on a common basis and makes predictions for new activators that may be discovered in the future.

TL;DR: Fundamental principles governing the behavior of macromolecules in a centrifugal field are summarized, and the application of these principles to the interpretation of data obtained from each type of experiment is reviewed.

TL;DR: These results demonstrate specific regions undergoing programmed cell death in normal development and increased LysoTracker staining in embryos exposed to hydroxyurea.

TL;DR: Methods for introducing fluorophores in RNA molecules are summarized here and are robust and readily applicable to the labeling of other types of probes.

TL;DR: The substance YC-1 has been shown to activate the enzyme independent of NO, to potentiate the effect of submaximally effective NO concentrations, and to turn carbon monoxide into an effective activator of soluble guanylyl cyclase.

TL;DR: Using multichannel confocal imaging, tissue slices that are single-, double-, or triple-labeled can be imaged in the living state in two or three spatial dimensions as well as in time, providing a means for investigating the cell-cell interaction and dynamic behavior of microglia and other cell types in live brain tissues cultured under various physiological conditions.

TL;DR: Two different transcriptional pulsing methods based on the c-fos serum-inducible promoter and the tetracycline-regulated promoter systems are described and discussed in detail to better elucidate the mechanistic steps and regulation underlying differential and selective mRNA turnover in mammalian cells.

TL;DR: In this paper, the authors describe procedures for the preparation, affinity purification, and initial characterization of site-specific antisera to acetylated histones, which can be specific not only for individual histones but also for histone isoforms acetylating at particular lysine residues.

TL;DR: The mechanisms controlling duplication of the metazoan genome are only beginning to be understood as mentioned in this paper and it is still unclear what organization of DNA sequences constitutes a chromosomal origin of DNA replication, and the regulation of origin activity during the cell cycle has not been fully revealed.

TL;DR: The use of yeast expression systems for the overproduction of complex multipolypeptide replication factors in yeast provides for a readily accessible and inexpensive source of these factors in large quantities.

TL;DR: Several lines of biochemical evidence suggest that guanylyl cyclase activator proteins (GCAPs) bind constitutively to an intracellular domain of RetGCs, which is necessary not only for normal photoresponses but also for photoreceptor viability since mutations in RetGC and in GCAP causephotoreceptor degeneration.