丁香实验_LOGO
登录
提问
提问
我要登录
|免费注册
丁香实验推荐阅读
Methods to Assess Myc Function in Intestinal Homeostasis, Regeneration, and Tumorigenesis

Within the intestinal epithelium, c-Myc has been characterized as a target of β-catenin-TCF signalling (He et al., Science 281:1509–1512, 1998). Given the most commonly mutated tumor suppressor gene within colorectal cancer (CRC) is the APC (Adenomatous Polyposis Coli) gene, a negative re ...

丁香实验推荐阅读
Generation of a Tetracycline Regulated Mouse Model of MYC-Induced T-Cell Acute Lymphoblastic Leukemia

The tetracycline regulatory system provides a tractable strategy to interrogate the role of oncogenes in the initiation and maintenance of tumorigenesis through both spatial and temporal control of expression. This approach has several potential advantages over convention ...

丁香实验推荐阅读
Studying Mycs Role in Metabolism Regulation

The MYC oncogene encodes a master transcription factor, Myc, which regulates genes involved in ribosome biogenesis, lipid synthesis, nucleic acid synthesis, intermediary metabolism, and cell growth and proliferation. The genomics of Myc target genes has been well-established th ...

丁香实验推荐阅读
Investigating Myc-Dependent Translational Regulation in Normal and Cancer Cells

There is an increasing realization that a primary role for Myc in driving cellular growth and cell cycle progression relies on Myc’s ability to increase the rate of protein synthesis. Myc induces myriad changes in both global and specific mRNA translation. Herein, we present three assays that al ...

丁香实验推荐阅读
A High-Throughput siRNA Screening Platform to Identify MYC-Synthetic Lethal Genes as Candidate Therapeutic Targets

Targeted therapeutics toward specific genes and pathways represent the future of oncological treatments. However, several commonly activated oncogenes, such as MYC, have proven difficult to target by pharmacological agents. To broaden the menu of potentially druggable thera ...

丁香实验推荐阅读
Genome-Wide Analysis of c-MYC-Regulated mRNAs and miRNAs, and c-MYC DNA Binding by Next-Generation Sequencing

The c-MYC oncogene is activated in ~50 % of all tumors, and its product, the c-MYC transcription factor, regulates numerous processes, which contribute to tumor initiation and progression. Therefore, the genome-wide characterization of c-MYC targets and their role in different tumor ent ...

丁香实验推荐阅读
Methods to Quantify microRNAs in the Myc Gene Network for Posttranscriptional Gene Repression

As a global transcription factor, Myc regulates both protein-coding genes and noncoding microRNA genes. Myc-activated or repressed miRNAs are involved in various pathways to affect tumorigenesis, mediate apoptosis, proliferation, angiogenesis, metastasis, and metaboli ...

丁香实验推荐阅读
Optimizing E. coli-Based Membrane Protein Production Using Lemo21(DE3) and GFP-Fusions

Optimizing the conditions for the overexpression of membrane proteins in E. coli and their subsequent purification is usually a laborious and time-consuming process. Combining the Lemo21(DE3) strain, which conveniently allows to identify the optimal expression intensity of a mem ...

丁香实验推荐阅读
Quantification of Protein Complexes by Blue Native Electrophoresis

Blue native electrophoresis (BNE) is a long established method for the analysis of native protein complexes. Applications of BNE range from investigating subunit composition, stoichiometry, and assembly of single protein complexes to profiling of whole complexomes. BNE is an indi ...

丁香实验推荐阅读
Single Channel Analysis of Membrane Proteins in Artificial Bilayer Membranes

The planar lipid bilayer technique is a powerful experimental approach for electrical single channel recordings of pore-forming membrane proteins in a chemically well-defined and easily modifiable environment. Here we provide a general survey of the basic materials and procedu ...

丁香实验推荐阅读
Reconstitution of Mitochondrial Presequence Translocase into Proteoliposomes

The isolation and functional reconstitution of large membrane protein complexes is an important step towards the biochemical characterization of such sophisticated molecular machines. Reconstitution is a multistep process that requires the mild solubilization of membr ...

丁香实验推荐阅读
Methods to Study the Biogenesis of Membrane Proteins in Yeast Mitochondria

The biogenesis of mitochondrial membrane proteins is an intricate process that relies on the import and submitochondrial sorting of nuclear-encoded preproteins and on the synthesis of mitochondrial translation products in the matrix. Subsequently, these polypeptides need to ...

丁香实验推荐阅读
An Assay to Monitor the Membrane Integration of Single-Span Proteins

In vitro import experiments with isolated organelles are a powerful tool for investigation of the biogenesis of proteins. A key issue in such experiments is an assay to distinguish between correctly and incorrectly imported proteins. Here we describe an assay to monitor in vitro the proper me ...

丁香实验推荐阅读
Analysis of Protein Translocation into the Endoplasmic Reticulum of Human Cells

The development of small-interfering RNA (siRNA)-mediated gene-silencing strategies has made it possible to study the transport of precursors of soluble and membrane proteins into the endoplasmic reticulum (ER) of human cells. In these approaches, a certain target gene is silenced in ...

丁香实验推荐阅读
Analyzing Membrane Dynamics with Live Cell Fluorescence Microscopy with a Focus on Yeast Mitochondria

With the availability of increasing numbers of fluorescent protein variants and state-of-the-art imaging techniques, live cell microscopy has become a standard procedure in modern cell biology. Fluorescent markers are used to visualize the dynamic processes that take place in liv ...

丁香实验推荐阅读
Membrane Partitioning and Translocation Studied by Isothermal Titration Calorimetry

The ability to bind to and translocate across lipid bilayers is of paramount importance for the extracellular administration of intracellularly active compounds in cell biology, medicinal chemistry, and drug development. A combination of the so-called uptake and release experim ...

丁香实验推荐阅读
Sedimentation Velocity Analytical Ultracentrifugation in Hydrogenated and Deuterated Solvents for the Characterization of Membrane Proteins

This chapter is a step-by-step protocol for setting up, realizing, and analyzing sedimentation velocity experiments in hydrogenated and deuterated solvents, in the context of the characterization of membrane protein, in terms of homogeneity, association state, and amount of bound d ...

丁香实验推荐阅读
Analyses of ProteinProtein Interactions by In Vivo Photocrosslinking in Budding Yeast

Recent development of methods for genetic incorporation of unnatural amino acids into proteins in live cells enables us to analyze protein interactions by site-specific photocrosslinking. Here we describe a method to incorporate p-benzoyl-l-phenylalanine (pBpa), a photorea ...

丁香实验推荐阅读
Scanning Fluorescence Correlation Spectroscopy in Model Membrane Systems

Fluorescence correlation spectroscopy (FCS) is an emerging technique employed in biophysical studies that exploits the temporal autocorrelation of fluorescence intensity fluctuations measured in a tiny volume (in the order of fL). The autocorrelation curve derived from the f ...

丁香实验推荐阅读
Peptide Interaction with and Insertion into Membranes

Natural and synthetic membrane active peptides as well as fragments from membrane proteins interact with membranes. In several cases, such interactions cause the insertion of the peptides to the membrane and their assembly within the lipid bilayer. Here we present spectroscopic appr ...

提问
扫一扫
丁香实验小程序二维码
实验小助手
丁香实验公众号二维码
关注公众号
反馈
TOP
打开小程序