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Image Credit: BioProtoc.3836

生物化学

Native Co-immunoprecipitation Assay to Identify Interacting Partners of Chromatin-associated Proteins in Mammalian Cells 天然共免疫沉淀法鉴定哺乳动物细胞中染色质相关蛋白的“伙伴蛋白质” 作者:Afzal Husain, Nasim A. Begum, Maki Kobayashi and Tasuku Honjo日期:12/05/2020,浏览量:2388,Q&A: 0

Protein-protein interactions play key roles in nuclear processes including transcription, replication, DNA damage repair, and recombination. Co-immunoprecipitation (Co-IP) followed by western blot or mass spectrometry is an invaluable approach to identify protein-protein interactions. One of the challenges in the Co-IP of a protein localized to

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生物物理学

Mechanical Characterization of Glandular Acini Using a Micro-indentation Instrument 使用微压痕仪对腺泡进行机械表征 作者:Christopher S. O’Bryan, Qiao Zhang, Tanmay P. Lele and Thomas E. Angelini日期:12/05/2020,浏览量:2091,Q&A: 0

The linker of nucleoskeleton and cytoskeleton (LINC) complex is responsible for tethering the nucleus to the cytoskeleton, providing a pathway for the cell’s nucleus to sense mechanical signals from the environment. Recently, we explored the role of the LINC complex in the development of glandular epithelial acini, such as those found in

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癌症生物学

A 3D Skin Melanoma Spheroid-Based Model to Assess Tumor-Immune Cell Interactions 基于球体的三维皮肤黑色素瘤模型评价肿瘤免疫细胞相互作用 作者:Marek Wagner and Shigeo Koyasu日期:12/05/2020,浏览量:1562,Q&A: 0
Three-dimensional (3D) tumor spheroids have the potential to bridge the gap between two-dimensional (2D) monolayer tumor cell cultures and solid tumors with which they share a significant degree of similarity. However, the progression of solid tumors is often influenced by the dynamic and reciprocal interactions between tumor and immune cells. ...
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发育生物学

Treadmill Running of Mouse as a Model for Studying Influence of Maternal Exercise on Offspring 以跑步机运动小鼠为模型研究母体运动对后代的影响 作者:Song Ah Chae, Jun Seok Son, Mei-Jun Zhu, Jeanene M. de Avila and Min Du日期:12/05/2020,浏览量:1523,Q&A: 0

Epidemiological studies robustly show the beneficial effects of maternal exercise in reducing maternal birth complications and improving neonatal outcomes, though underlying mechanisms remain poorly understood. To facilitate mechanistic exploration, a protocol for maternal exercise of mice is established, with the regimen following the exercise

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免疫学

Analysis of B Cell Migration by Intravital Microscopy B细胞迁移的活体成像仪观察分析 作者:Michael Schnoor, Leopoldo Santos-Argumedo, Daniel Alberto Girón-Pérez and Eduardo Vadillo日期:12/05/2020,浏览量:1528,Q&A: 0

During immune responses, B cells home to lymph nodes (LN), where they encounter antigens. Homing starts with capture and L-selectin-dependent rolling on the activated endothelium of high endothelial venules (HEV). After recognition of chemokines presented on HEV, activation of B cell integrins occurs mediating firm arrest. Subsequently, B cells

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Protocol for Isolation, Stimulation and Functional Profiling of Primary and iPSC-derived Human NK Cells 原代和iPSC来源的人类NK细胞的分离、刺激和功能分析方案 作者:Janine E. Melsen, Maria Themeli, Monique M. van Ostaijen-ten Dam, Els van Beelen, Gertjan Lugthart, Rob C. Hoeben, Marco W. Schilham and Harald M. Mikkers日期:12/05/2020,浏览量:3115,Q&A: 0

Natural killer (NK) cells are innate immune cells, characterized by their cytotoxic capacity, and chemokine and cytokine secretion upon activation. Human NK cells are identified by CD56 expression. Circulating NK cells can be further subdivided into the CD56bright (~10%) and CD56dim NK cell subsets (~90%). NK cell-like cells can also be derived

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微生物学

Ribosome Purification from an α-proteobacterium and rRNA Analysis by Northern Blot α-变形杆菌核糖体的纯化及Northern印迹分析 作者:Liang Yin and Caroline S. Harwood日期:12/05/2020,浏览量:1043,Q&A: 0
Ribosomes are an integral part of cellular life. They are complex molecular machines consisting of multiple ribosomal proteins and RNAs. To study different aspects of ribosome composition, many methods have been developed over the decades. Here, we describe how to purify ribosomes from the α-proteobacterium Rhodopseudomonas palustris ...
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Combining Gel Retardation and Footprinting to Determine Protein-DNA Interactions of Specific and/or Less Stable Complexes 结合凝胶阻滞和足印法测定特异性和/或不稳定复合物的蛋白质-DNA相互作用 作者:Meng-Lun Hsieh, Alice Boulanger, Leslie G. Knipling and Deborah M. Hinton日期:12/05/2020,浏览量:1058,Q&A: 0

DNA footprinting is a classic technique to investigate protein-DNA interactions. However, traditional footprinting protocols can be unsuccessful or difficult to interpret if the binding of the protein to the DNA is weak, the protein has a fast off-rate, or if several different protein-DNA complexes are formed. Our protocol differs from traditional

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Expression and Purification of Recombinant Skd3 (Human ClpB) Protein and Tobacco Etch Virus (TEV) Protease from Escherichia coli 重组Skd3(人ClpB)蛋白和烟草蚀刻病毒(TEV)蛋白酶在大肠杆菌中的表达与纯化 作者:Ryan R. Cupo and James Shorter日期:12/05/2020,浏览量:1825,Q&A: 0

Skd3 (encoded by human CLPB) is a mitochondrial AAA+ protein comprised of an N-terminal ankyrin-repeat domain and a C-terminal HCLR-clade nucleotide-binding domain. The function of Skd3 has long remained unknown due to challenges in purifying the protein to high quality and near homogeneity. Recently we described Skd3 as a human mitochondrial

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分子生物学

Charging State Analysis of Transfer RNA from an α-proteobacterium α-变形杆菌转运RNA的装载状态分析 作者:Liang Yin and Caroline S. Harwood日期:12/05/2020,浏览量:1069,Q&A: 0
Transfer RNA (tRNA) is an essential link between the genetic code and proteins. During the process of translation, tRNA is charged with its cognate amino acid and delivers it to the ribosome, thus serving as a substrate of protein synthesis. To analyze the charging state of a particular tRNA, total RNA is purified and analyzed on an acid-urea gel. ...
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神经科学

Using the Parafilm-assisted Microdissection (PAM) Method to Sample Rodent Nucleus Accumbens 应用膜辅助显微切割(PAM)对啮齿类伏隔核进行采样 作者:Marco Pagliusi Jr., Arthur F. Brandão, Gabriel G. Zanetti, Ivan J. M. Bonet, Cesar R. Sartori and Andre S. Vieira日期:12/05/2020,浏览量:1088,Q&A: 0
Microdissection techniques are very important for anatomical and functional studies focused on neuroscience, where it is often necessary microdissect specific brain areas to perform molecular or anatomical analyses. The parafilm®-assisted microdissection (PAM) was previously described and involves the microdissection of tissue ...
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Headpost Surgery for in vivo Electrophysiological Recording in the Mouse Inferior Colliculus during Locomotion 用于运动过程中小鼠下丘体内电生理记录的头部手术 作者:Yoonsun Yang and Gunsoo Kim日期:12/05/2020,浏览量:1179,Q&A: 0

The inferior colliculus (IC) is a critical midbrain integration center for auditory and non-auditory information. Although much is known about the response properties of the IC neurons to auditory stimuli, how the IC neural circuits function during movement such as locomotion remains poorly understood. Mice offer a valuable model in this respect,

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Use of Optogenetic Amyloid-β to Monitor Protein Aggregation in Drosophila melanogaster, Danio rerio and Caenorhabditis elegans 利用光遗传学淀粉样蛋白-β监测黑腹果蝇、斑马鱼和秀丽隐杆线虫体内蛋白质聚集 作者:Prameet Kaur, Caroline Kibat, Emelyne Teo, Jan Gruber, Ajay Mathuru and Nicholas S. Tolwinski日期:12/05/2020,浏览量:2030,Q&A: 0

Alzheimer’s Disease (AD) has long been associated with accumulation of extracellular amyloid plaques (Aβ) originating from the Amyloid Precursor Protein. Plaques have, however, been discovered in healthy individuals and not all AD brains show plaques, suggesting that extracellular Aβ aggregates may play a smaller role than

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植物科学

Chromatin Immunoprecipitation (ChIP) to Assess Histone Marks in Auxin-treated Arabidopsis thaliana Inflorescence Tissue 染色质免疫沉淀法(ChIP)检测生长素处理拟南芥花序组织中的组蛋白标记 作者:André Kuhn and Lars Østergaard日期:12/05/2020,浏览量:1621,Q&A: 0
Chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) or high-throughput sequencing (ChIP-seq) has become the gold standard for the identification of binding sites of DNA binding proteins and the localization of histone modification on a locus-specific or genome-wide scale, respectively. ChIP experiments can be divided into seven ...
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Pipecolic Acid Quantification Using Gas Chromatography-coupled Mass Spectrometry 哌啶酸的气相色谱-质谱定量分析 作者:Keshun Yu, Huazhen Liu and Pradeep Kachroo日期:12/05/2020,浏览量:1519,Q&A: 0

Pipecolic acid (Pip), a non-proteinacious product of lysine catabolism, is an important regulator of immunity in plants and humans alike. For instance, Pip accumulation is associated with the genetic disorder Zellweger syndrome, chronic liver diseases, and pyridoxine-dependent epilepsy in humans. In plants, Pip accumulates upon pathogen infection

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