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Image Credit: BioProtoc.3249

生物化学

Glucosidase Inhibition to Study Calnexin-assisted Glycoprotein Folding in Cells 利用葡萄糖苷酶抑制作用研究细胞中钙连蛋白辅助糖蛋白折叠 作者:Hao Wang and Qingyu Wu日期:06/05/2019,浏览量:2984,Q&A: 0
Calnexin is a chaperone protein that plays a critical role in glycoprotein folding in the endoplasmic reticulum (ER). The function of calnexin depends on its binding to monoglucosylated oligosaccharides on nascent glycoproteins, whereas the generation of monoglucosylated oligosaccharides depends on the activity of α-glucosidases I and II, which ...
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Tryptophan Fluorescence Quenching Assays for Measuring Protein-ligand Binding Affinities: Principles and a Practical Guide 用于检测蛋白配体结合亲和力的色氨酸荧光猝灭试验:原理和操作指南 作者:Anthony Yammine, Jinlong Gao and Ann H. Kwan日期:06/05/2019,浏览量:13265,Q&A: 0
Tryptophan fluorescence quenching is a type of fluorescence spectroscopy used for binding assays. The assay relies on the ability to quench the intrinsic fluorescence of tryptophan residues within a protein that results from changes in the local environment polarity experienced by the tryptophan(s) upon the addition of a binding partner or ligand. ...
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Cross-linking, Immunoprecipitation and Proteomic Analysis to Identify Interacting Proteins in Cultured Cells 利用交联、免疫沉淀和蛋白质组学分析法鉴定培养细胞中互作的蛋白质 作者:Hao Wang, Meiling He, Belinda Willard and Qingyu Wu日期:06/05/2019,浏览量:9182,Q&A: 1
Extracellular expression is essential for the function of secreted and cell surface proteins. Proper intracellular trafficking depends on protein interactions in multiple subcellular compartments. Co-immunoprecipitation and the yeast two-hybrid system are commonly used to investigate protein-protein interactions. These methods, however, depend on ...
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A Detailed Protocol for Large-scale Recombinant Expression and Validation of Human FGFR2 with Its Transmembrane and Extracellular Domains in Escherichia coli 人FGFR2跨膜区域和胞外区域在大肠杆菌中的大规模重组表达和验证的详细方法 作者:Adam Bajinting and Ho Leung Ng日期:06/05/2019,浏览量:3491,Q&A: 0
Receptor tyrosine kinases (RTKs) are an important class of transmembrane receptors that mediate some of the most crucial biochemical pathways essential to the growth, differentiation, and survival of a cell and thus, are highly involved in cancers. Due to the complexity of RTKs having biochemically different domains including a transmembrane ...
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免疫学

Detection of Antigen-specific T cells in Spleens of Vaccinated Mice Applying 3[H]-Thymidine Incorporation Assay and Luminex Multiple Cytokine Analysis Technology 利用3[H]-胸苷掺入分析和Luminex多重细胞因子分析技术检测免疫小鼠脾脏中抗原特异性T细胞 作者:Maria Agallou and Evdokia Karagouni日期:06/05/2019,浏览量:7471,Q&A: 0
For many infectious diseases T cells are an important part of naturally acquired protective immune responses, and inducing these by vaccination has been the aim of much research. Here, we describe a protocol for the analysis of vaccine-induced antigen-specific immune responses. For this purpose, cells of whole spleens obtained from vaccinated ...
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Measurement of CD74 N-terminal Fragment Accumulation in Cells Treated with SPPL2a Inhibitor SPPL2a抑制剂处理细胞后的CD74N端片段聚集的检测 作者:Rubén Martínez-Barricarte, Xiao-Fei Kong and Jean-Laurent Casanova日期:06/05/2019,浏览量:3536,Q&A: 0
The recent discovery of human signal peptide peptidase-like 2a (SPPL2a) deficiency in humans revealed the toxicity associated with the accumulation of one of its substrates, CD74 N-terminal fragment (CD74-NTF), for certain type of dendritic cells (cDC2). We developed a two-step protocol for monitoring the accumulation of this molecule in different ...
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分子生物学

Improved Mammalian Mitochondrial RNA Isolation 一种改进的哺乳动物线粒体RNA分离方法 作者:Jinliang Huang and Geng Wang日期:06/05/2019,浏览量:4711,Q&A: 0
Mitochondria have two sets of RNAs. One is encoded in mitochondrial genome, and the other that consists of imported RNAs within mitochondria and cytosolic RNAs associated with mitochondrial outer membrane is encoded in the nucleus. These mitochondrial RNAs play important roles in mitochondrion biosynthesis and signaling in and out of mitochondria. ...
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Minigene Assay to Evaluate CRISPR/Cas9-based Excision of Intronic Mutations that Cause Aberrant Splicing in Human Cells Minigene技术评估会导致人细胞产生异常剪切的基于CRISPR/Cas9 的内含子突变切除 作者:David J. Sanz and Patrick T. Harrison日期:06/05/2019,浏览量:3769,Q&A: 0
The construction of Hybrid minigenes provides a robust and simple strategy to study the effects of disease-causing mutations on mRNA splicing when biological material from patient cells is not available. Hybrid minigenes can be used as splicing reporter plasmids allow RNA expression and heterologous splicing reactions between synthetic splicing ...
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Organelle-associated rRNA Degradation 附着在细胞器上的rRNA降解 作者:Jinliang Huang and Geng Wang日期:06/05/2019,浏览量:3431,Q&A: 0
Cytosolic rRNAs are highly dynamic and can be degraded under conditions such as apoptosis, starvation and magnesium depletion. The degradation is also related to their specific localization, as fractions of cytosolic ribosomes are localized on the surfaces of intracellular organelles, such as endoplasmic reticulum (ER) and mitochondria. Such ...
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神经科学

Accessing Olfactory Habituation in Drosophila melanogaster with a T-maze Paradigm 使用T迷宫范例模拟黑腹果蝇的嗅觉习性 作者:Ourania Semelidou, Summer F. Acevedo and Efthimios M.C. Skoulakis日期:06/05/2019,浏览量:3934,Q&A: 0
Habituation is the process whereby perceptual changes alter the value of environmental stimuli, enabling salience filtering. This behavioral response decrement is a form of non-associative learning, where the subject learns about the stimulus and does not involve sensory adaptation, sensory or motor fatigue. The range of behavioral responses in D. ...
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植物科学

Transient Expression Assay in Strawberry Fruits 草莓果实中的瞬时表达检测 作者:Mengting Pi, Qi Gao and Chunying Kang日期:06/05/2019,浏览量:4718,Q&A: 0
Strawberry, including the woodland strawberry Fragaria vesca (2x) and the cultivated strawberry (Fragaria × ananassa, 8x), has emerged as a model system for studying fruit development and ripening. Transient expression provides a quick assay for gene functions or gene interactions. In strawberry, ...
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Simple Method to Determine Protein Redox State in Arabidopsis thaliana 一种确定拟南芥蛋白氧化还原状态的简单方法 作者:Keisuke Yoshida and Toru Hisabori日期:06/05/2019,浏览量:4272,Q&A: 1
Thiol-based redox regulation is a posttranslational protein modification that plays a key role in many biological aspects. To understand its regulatory functions, we need a method to directly assess protein redox state in vivo. Here we present a simple procedure to determine protein redox state in a model plant Arabidopsis thaliana ...
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