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丁香实验推荐阅读
Genome-Wide RNAi Screening to Identify Regulators of Oncogene-Induced Cellular Senescence

RNA interference (RNAi) is a powerful research tool that can be used to turn off—or silence—the expression of a specific gene. In recent years, RNAi screening on a genome-wide scale has provided the opportunity to identify factors and pathways involved in complex biological processes in a syste ...

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Profiling the Metabolic Signature of Senescence

Aging is a complex process, which involves changes in different cellular functions that all can be integrated on the metabolite level. This means that different gene regulation pathways that affect aging might lead to similar changes in metabolism and result in a metabolic signature of sene ...

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Monitoring Nutrient Signaling Through the Longevity Protein p66SHC1

Nutrient availability and nutrient-dependent biochemical signals represent major determinants of cellular senescence and organismal aging. The present chapter describes simple procedures to reliably evaluate the response of cultured cell to nutrients through the long ...

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Methods to Investigate the Role of SIRT1 in Endothelial Senescence

Sirtuins are a family of proteins with NAD+-dependent deacetylase or mono-ADP-ribosyltransferase activity. SIRT1, the mammalian ortholog most closely related to Sir2 (the first gene of this family discovered in yeast), exhibits anti-senescence activity in a wide range of mammalian ...

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Monitoring Oncogenic B-RAF-Induced Senescence in Melanocytes

The B-RAF kinase is a downstream effector of the RAS family of proto-oncogenes and is constitutively activated in the majority of human melanomas. The common oncogenic B-RAFV600E mutant cooperates with additional genetic lesions to transform immortal murine and human cells. In primary ...

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Raman Spectroscopy for the Detection of AGEs/ALEs

Raman spectroscopy is a noninvasive, nondestructive tool for capturing multiplexed biochemical information across diverse molecular species including proteins, lipids, DNA, and mineralizations. Based on light scattering from molecules, cells, and tissues, it is possible to ...

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Cell Senescence as Both a Dynamic and a Static Phenotype

It has been 50 years since cellular senescence was first described in human diploid fibroblasts (HDFs), yet its mechanism as well as its physiological and clinical implications are still not fully appreciated. Recent progress suggests that cellular senescence is a collective phenoty ...

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Assays for the Measurement of Lipid Peroxidation

Physical and emotional stress, metabolic alterations, carcinogenesis or inflammation are conditions that can trigger oxidative stress, which is defined as a balance shift of redox reactions towards oxidation, resulting in the increase of reactive oxygen species (ROS). ROS are cont ...

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Quantification of Protein Carbonylation

Protein carbonylation is the most commonly used measure of oxidative modification of proteins. It is most often measured spectrophotometrically or immunochemically by derivatizing proteins with the classical carbonyl reagent 2,4 dinitrophenylhydrazine (DNPH). We prese ...

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Measuring Reactive Oxygen Species in Senescent Cells

Countless studies have implicated reactive oxygen species (ROS) and mitochondrial dysfunction in the ageing process. During cellular senescence, the ultimate and irreversible loss of replicative capacity of somatic cells grown in culture, several studies have reported incre ...

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Detection of Nuclear Envelope Alterations in Senescence

Gene mutations that cause defects in the nuclear envelope are responsible for progeroid syndromes, characterized by exacerbated cell senescence and accelerated aging. Consequently, morphological abnormalities of the nucleus represent a cellular phenotype whose analys ...

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Methods for the Assessment of Telomere Status

Most methods for examining telomere functionality have relied on measurements of telomeric DNA by hybridization or quantitative PCR. While these techniques yield measures of telomeric DNA length, they generate whole-population results. However, telomeric DNA lengths on diff ...

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Assessment and Quantification of Telomerase Enzyme Activity

The enzyme telomerase is activated in 80–90% of all human malignancies and immortal cell lines, where it functions to maintain the integrity of chromosomal-end structures called telomeres. Telomerase enzyme activity can be detected in whole cell lysates by a polymerase chain reaction ( ...

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Monitoring DNA Damage During Cell Senescence

Cellular senescence is a state of irreversible cell cycle arrest, accompanied by and in most cases driven by a persistent DNA damage response (DDR), which may be activated by uncapped telomeres or other forms of DNA damage. DNA damage foci, therefore, are an important part of the signaling pathway th ...

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Detection of Senescence-Associated Heterochromatin Foci (SAHF)

One of the most prominent features of cellular senescence, a stress response that prevents the propagation of cells that have accumulated potentially oncogenic alterations, is a permanent loss of proliferative potential. Thus, at odds with quiescent cells, which resume proliferat ...

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Unbiased Characterization of the Senescence-Associated Secretome Using SILAC-Based Quantitative Proteomics

Approaches based on the combination of mass spectrometry (MS) and quantitative methods have the potential to generate unbiased, thorough proteomic catalogues. In particular, stable isotope labeling with amino acid in cell culture (SILAC) has been used to perform highly accurate qua ...

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Detection of the Senescence-Associated Secretory Phenotype (SASP)

Cellular senescence suppresses cancer by eliminating potentially oncogenic cells, participates in tissue repair, contributes to cancer therapy, and promotes organismal aging. Numerous activities of senescent cells depend on the aptitude of these cells to secrete myriads of bi ...

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Simultaneous Cloning and Selection of Hybridomas and Transfected Cell Lines in Semisolid Media

Selection and cloning are essential but often laborious and time-consuming steps during the generation of hybridomas and genetically modified cell lines that produce monoclonal antibodies or other proteins with desired properties. Methods for the simultaneous selection and c ...

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In Vitro Generation of Human T Regulatory Cells: Generation, Culture, and Analysis of FOXP3-Transduced T Cells

T regulatory cells (Tregs) suppress immune responses and therefore have potential to be used in the clinic as a cellular therapy for autoimmune disease and to prevent rejection of transplanted organs. Obtaining sufficient numbers of these cells for therapeutic use is a challenge, however, ...

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Generation, Isolation, and Engraftment of In Vitro-Derived Human T Cell Progenitors

T cells typically differentiate via a series of coordinated steps within the highly specialized microenvironment of the thymus. Traditionally, human T-lymphopoiesis in vitro has been studied using the hybrid human/mouse fetal thymic organ culture system. Pioneering work by McCu ...

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