• Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Biochemistry

The Long-lived Protein Degradation Assay: an Efficient Method for Quantitative Determination of the Autophagic Flux of Endogenous Proteins in Adherent Cell Lines Authors:  Morten Luhr, Frank Sætre and Nikolai Engedal, date: 05/05/2018, view: 1786, 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 ...
More >>

Cancer Biology

An Image-based Assay for High-throughput Analysis of Cell Proliferation and Cell Death of Adherent Cells Authors:  Paula Szalai and Nikolai Engedal, date: 05/05/2018, view: 1603, 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. ...
More >>

Cell Biology

Quantifying Podocytes and Parietal Epithelial Cells in Human Urine Using Liquid-based Cytology and WT1 Immunoenzyme Staining Authors:  Hiroyuki Ohsaki, Toru Matsunaga, Taishi Fujita, Yasunori Tokuhara, Shingo Kamoshida and Tadashi Sofue, date: 05/05/2018, view: 1295, 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.
More >>

Immunology

Purification of Total RNA from DSS-treated Murine Tissue via Lithium Chloride Precipitation Authors:  Emilie Viennois, Anika Tahsin and Didier Merlin, date: 05/05/2018, view: 1383, 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.
More >>
Nab Escaping AAV Mutants Isolated from Mouse Muscles Authors:  Zheng Chai, R. Jude Samulski and Chengwen Li, date: 05/05/2018, view: 1278, 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 ...
More >>

Microbiology

Glycogen and Extracellular Glucose Estimation from Cyanobacteria Synechocystis sp. PCC 6803 Authors:  Md. Rezaul Islam Khan, Yushu Wang, Shajia Afrin, Lin He and Gang Ma, date: 05/05/2018, view: 1058, 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 ...
More >>
A Modified Low-quantity RNA-Seq Method for Microbial Community and Diversity Analysis Using Small Subunit Ribosomal RNA Authors:  Yong-Wei Yan, Ting Zhu, Bin Zou and Zhe-Xue Quan, date: 05/05/2018, view: 821, 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 ...
More >>
Time-of-addition and Temperature-shift Assays to Determine Particular Step(s) in the Viral Life Cycle that is Blocked by Antiviral Substance(s) Authors:  Chie Aoki-Utsubo, Ming Chen and Hak Hotta, date: 05/05/2018, view: 1047, 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 ...
More >>
Assessing the Efficacy of Small Molecule Inhibitors in a Mouse Model of Persistent Norovirus Infection Authors:  Jana Van Dycke, Johan Neyts and Joana Rocha-Pereira, date: 05/05/2018, view: 1216, 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 ...
More >>
Infection Process Observation of Magnaporthe oryzae on Barley Leaves Author:  Xiao-Lin Chen, date: 05/05/2018, view: 1519, 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 ...
More >>
Detecting the Interaction of Double-stranded RNA Binding Protein, Viral Protein and Primary miRNA Transcript by Co-immunoprecipitation in planta Authors:  Lijia Zheng, Chao Zhang, Jianguo Wu and Yi Li, date: 05/05/2018, view: 825, 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 ...
More >>
Heterologous Expression and Purification of the CRISPR-Cas12a/Cpf1 Protein Authors:  Prarthana Mohanraju, John van der Oost, Martin Jinek and Daan C. Swarts, date: 05/05/2018, view: 2223, 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 ...
More >>

Molecular Biology

Mammalian Cell-derived Vesicles for the Isolation of Organelle Specific Transmembrane Proteins to Conduct Single Molecule Studies Authors:  Faruk H. Moonschi, Ashley M. Fox-Loe, Xu Fu and Chris I. Richards, date: 05/05/2018, view: 1198, 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 ...
More >>

Plant Science

Evaluation of the Condition of Respiration and Photosynthesis by Measuring Chlorophyll Fluorescence in Cyanobacteria Authors:  Takako Ogawa and Kintake Sonoike , date: 05/05/2018, view: 923, 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 ...
More >>
Functional Evaluation of the Signal Peptides of Secreted Proteins Authors:  Weixiao Yin, Yufu Wang, Tao Chen, Yang Lin and Chaoxi Luo, date: 05/05/2018, view: 1069, Q&A: 0
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 ...
More >>

Stem Cell

Cobblestone Area-forming Cell Assay of Mouse Bone Marrow Hematopoietic Stem Cells Authors:  Surya Amarachintha and Qishen Pang, date: 05/05/2018, view: 1456, 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 ...
More >>
We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.