• 卷册 11, 2021
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Image Credit: BioProtoc.2827

生物化学

The Long-lived Protein Degradation Assay: an Efficient Method for Quantitative Determination of the Autophagic Flux of Endogenous Proteins in Adherent Cell Lines 长寿蛋白降解测定法:定量测定粘附细胞系中内源性蛋白自噬通量的有效方法 作者:Morten Luhr, Frank Sætre and Nikolai Engedal日期:05/05/2018,浏览量:8011,Q&A: 0
Autophagy is a key player in the maintenance of cellular homeostasis in eukaryotes, and numerous diseases, including cancer and neurodegenerative disorders, are associated with alterations in autophagy. The interest for studying autophagy has grown intensely in the last two decades, and so has the arsenal of methods utilised to study this highly ...
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癌症生物学

An Image-based Assay for High-throughput Analysis of Cell Proliferation and Cell Death of Adherent Cells 采用基于成像的分析法进行粘附细胞的细胞增殖和细胞死亡高通量分析 作者:Paula Szalai and Nikolai Engedal日期:05/05/2018,浏览量:7591,Q&A: 0
In this protocol, we describe a method to monitor cell proliferation and death by live-cell imaging of propidium iodide (PI)-stained adherent mammalian cells. PI is widely used to assess cell death. However, it is usually used in end-point assays. Recently, we implemented the use of PI for real-time cell death assessment by automated imaging. ...
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细胞生物学

Quantifying Podocytes and Parietal Epithelial Cells in Human Urine Using Liquid-based Cytology and WT1 Immunoenzyme Staining 使用基于液体的细胞学检查和WT1免疫酶染色定量人尿中的足细胞和壁层上皮细胞 作者:Hiroyuki Ohsaki, Toru Matsunaga, Taishi Fujita, Yasunori Tokuhara, Shingo Kamoshida and Tadashi Sofue日期:05/05/2018,浏览量:5488,Q&A: 0
In glomerular disease, podocytes and parietal epithelial cells (PECs) are shed in the urine. Therefore, urinary podocytes and PECs are noninvasive biomarkers of glomerular disease. The purpose of this protocol is to employ immunocytochemistry to detect podocytes and PECs, using the WT1 antibody on liquid-based cytology slides.
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免疫学

Purification of Total RNA from DSS-treated Murine Tissue via Lithium Chloride Precipitation 通过氯化锂沉淀法纯化DSS处理的鼠组织的总RNA 作者:Emilie Viennois, Anika Tahsin and Didier Merlin日期:05/05/2018,浏览量:6023,Q&A: 0
We have developed a protocol to purify RNA from DSS (Dextran Sulfate Sodium)-treated mouse tissues. This method, which prevents downstream inhibition of q-RT-PCR observed in DSS-treated tissues, relies on successive precipitations with lithium chloride.
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Nab Escaping AAV Mutants Isolated from Mouse Muscles 从小鼠肌肉中分离Nab逃脱AAV突变体 作者:Zheng Chai, R. Jude Samulski and Chengwen Li日期:05/05/2018,浏览量:5145,Q&A: 0
Neutralizing antibodies (Nabs) are a major challenge in clinical trials of adeno-associated virus (AAV) vector gene therapy, because Nabs are able to inhibit AAV transduction in patients. We have successfully isolated several novel Nab-escaped AAV chimeric capsids in mice by administrating a mixture of AAV shuffled library and patient serum. These ...
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微生物学

Glycogen and Extracellular Glucose Estimation from Cyanobacteria Synechocystis sp. PCC 6803 蓝藻集胞藻PCC 6803中糖原和胞外葡萄糖估测 作者:Md. Rezaul Islam Khan, Yushu Wang, Shajia Afrin, Lin He and Gang Ma日期:05/05/2018,浏览量:4850,Q&A: 0
Cyanobacteria, which have the extraordinary ability to grow using sunlight and carbon dioxide, are emerging as a green host to produce value-added products. Exploitation of this highly promising host to make products may depend on the ability to modulate the glucose metabolic pathway; it is the key metabolic pathway that generates intermediates ...
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A Modified Low-quantity RNA-Seq Method for Microbial Community and Diversity Analysis Using Small Subunit Ribosomal RNA 改良的低量 RNA-Seq法通过检测小亚基核糖体RNA以分析微生物群落和多样性 作者:Yong-Wei Yan, Ting Zhu, Bin Zou and Zhe-Xue Quan日期:05/05/2018,浏览量:3950,Q&A: 0
We propose a modified RNA-Seq method for small subunit ribosomal RNA (SSU rRNA)-based microbial community analysis that depends on the direct ligation of a 5’ adaptor to RNA before reverse-transcription. The method requires only a low-input quantity of RNA (10-100 ng) and does not require a DNA removal step. Using this method, we could obtain more ...
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Time-of-addition and Temperature-shift Assays to Determine Particular Step(s) in the Viral Life Cycle that is Blocked by Antiviral Substance(s) 改变药物添加时间及变温分析实验确定抗病毒物质所作用的病毒生命周期中的步骤 作者:Chie Aoki-Utsubo, Ming Chen and Hak Hotta日期:05/05/2018,浏览量:6533,Q&A: 0
Viruses infect their host cells to produce progeny virus particles through the sequential steps of the viral life cycle, such as viral attachment, entry, penetration and post-entry events. This protocol describes time-of-addition and temperature-shift assays that are employed to explore which step(s) in the viral life cycle is blocked by an ...
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Assessing the Efficacy of Small Molecule Inhibitors in a Mouse Model of Persistent Norovirus Infection 小分子抑制剂在持续性诺如病毒感染小鼠模型中的疗效评估 作者:Jana Van Dycke, Johan Neyts and Joana Rocha-Pereira日期:05/05/2018,浏览量:4651,Q&A: 0
Human norovirus is the most common cause of acute gastroenteritis worldwide, resulting in estimated mortality of ~210,000 each year, of whom most are children under the age of five. However, norovirus can infect people of all age groups. There is a risk of prolonged infection in children, the elderly and patients who are immunocompromised. To ...
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Infection Process Observation of Magnaporthe oryzae on Barley Leaves 大麦叶片上稻瘟病菌感染过程观察 作者:Xiao-Lin Chen日期:05/05/2018,浏览量:6853,Q&A: 0
Rice blast and wheat blast caused by Magnaporthe oryzae is a serious threat to rice and wheat production. Appropriate methods for observing M. oryzae infection process are important for study of the fungal infection mechanisms, plant resistance reactions, and host-M. oryzae interactions. The rice leaf sheath is commonly ...
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Detecting the Interaction of Double-stranded RNA Binding Protein, Viral Protein and Primary miRNA Transcript by Co-immunoprecipitation in planta 通过免疫共沉淀法检测植物中双链RNA结合蛋白、病毒蛋白和初级miRNA转录产物的交互作用 作者:Lijia Zheng, Chao Zhang, Jianguo Wu and Yi Li日期:05/05/2018,浏览量:4293,Q&A: 0
MicroRNAs (miRNAs) play important roles in plant growth, development, and response to infection by microbes. Double-stranded RNA binding protein 1 (DRB1) facilitates the processing of primary miRNA transcripts into mature miRNAs. Recently, we found that NS3 protein encoded by rice stripe virus (RSV) associates with DRB1 and promotes miRNA ...
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Heterologous Expression and Purification of the CRISPR-Cas12a/Cpf1 Protein CRISPR-Cas12a/Cpf1的异源表达和纯化 作者:Prarthana Mohanraju, John van der Oost, Martin Jinek and Daan C. Swarts日期:05/05/2018,浏览量:14072,Q&A: 3
This protocol provides step by step instructions (Figure 1) for heterologous expression of Francisella novicida Cas12a (previously known as Cpf1) in Escherichia coli. It additionally includes a protocol for high-purity purification and briefly describes how activity assays can be performed. These protocols can also be used for ...
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分子生物学

Mammalian Cell-derived Vesicles for the Isolation of Organelle Specific Transmembrane Proteins to Conduct Single Molecule Studies 哺乳动物细胞源性囊泡用于分离细胞器特异性跨膜蛋白以进行单分子研究 作者:Faruk H. Moonschi, Ashley M. Fox-Loe, Xu Fu and Chris I. Richards日期:05/05/2018,浏览量:5062,Q&A: 0
Cell-derived vesicles facilitate the isolation of transmembrane proteins in their physiological membrane maintaining their structural and functional integrity. These vesicles can be generated from different cellular organelles producing, housing, or transporting the proteins. Combined with single-molecule imaging, isolated organelle specific ...
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植物科学

Evaluation of the Condition of Respiration and Photosynthesis by Measuring Chlorophyll Fluorescence in Cyanobacteria 通过测定蓝藻叶绿素荧光评估呼吸和光合作用的条件 作者:Takako Ogawa and Kintake Sonoike 日期:05/05/2018,浏览量:5647,Q&A: 0
Chlorophyll fluorescence measurements have been widely used to monitor the condition of photosynthesis. Furthermore, chlorophyll fluorescence from cyanobacteria reflects the condition of respiration, since cyanobacterial photosynthesis shares several components of electron transport chain with respiration. This protocol presents the method to ...
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Functional Evaluation of the Signal Peptides of Secreted Proteins 分泌蛋白信号肽功能评估 作者:Weixiao Yin, Yufu Wang, Tao Chen, Yang Lin and Chaoxi Luo日期:05/05/2018,浏览量:8107,Q&A: 4
Here, we describe a method that can be used to evaluate the function of predicted signal peptides. This method utilizes the yeast Saccharomyces cerevisiae YTK12 strain and pSUC2 vector in which the pSUC2 vector with fused predicted signal peptide is transformed into yeast. The function of the signal peptides can be evaluated by using ...
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干细胞

Cobblestone Area-forming Cell Assay of Mouse Bone Marrow Hematopoietic Stem Cells 小鼠骨髓造血干细胞的鹅卵石样区域形成细胞测定 作者:Surya Amarachintha and Qishen Pang日期:05/05/2018,浏览量:7504,Q&A: 0
Bone Marrow Hematopoietic Stem Cells (HSCs) require bone marrow microenvironment for their maintenance and proliferation. Culture of Bone Marrow Mesenchymal Stromal Cells (MSCs) provides appropriate environmental signals for HSCs survival in vitro. Here, we provide a detailed protocol that describes culture conditions for MSCs, flow ...
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