• Volume 10, 2020
  • Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Past Issues in 2020

Volume: 10 Issue: 4

Biochemistry

Photoactivable Cholesterol as a Tool to Study Interaction of Influenza Virus Hemagglutinin with Cholesterol Authors:  Bodan Hu, Mohamed Rasheed Gadalla, Christoph Thiele and Michael Veit, date: 02/20/2020, view: 937, Q&A: 0
Non-covalent binding of cholesterol to the transmembrane region of proteins affect their functionalities, but methods to prove such an interaction are rare. We describe our protocol to label the hemagglutinin (HA) of Influenza virus with a cholesterol derivative in living cells or with immunoprecipitated protein. We synthesized a “clickable” ...
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Cell Biology

ChIP-Seq from Limited Starting Material of K562 Cells and Drosophila Neuroblasts Using Tagmentation Assisted Fragmentation Approach Authors:  Junaid Akhtar, Piyush More and Steffen Albrecht, date: 02/20/2020, view: 637, Q&A: 0
Chromatin immunoprecipitation is extensively used to investigate the epigenetic profile and transcription factor binding sites in the genome. However, when the starting material is limited, the conventional ChIP-Seq approach cannot be implemented. This protocol describes a method that can be used to generate the chromatin profiles from as low as ...
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CRISPR-Cas9 Genome Editing of Plasmodium knowlesi Authors:  Franziska Mohring, Melissa N. Hart, Avnish Patel, David A. Baker and Robert W. Moon, date: 02/20/2020, view: 966, Q&A: 0
Plasmodium knowlesi is a zoonotic malaria parasite in Southeast Asia that can cause severe and fatal malaria in humans. The main hosts are Macaques, but modern diagnostic tools reveal increasing numbers of human infections. After P. falciparum, P. knowlesi is the only other malaria parasite capable of being maintained in ...
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Estimating Cellular Abundances of Halo-tagged Proteins in Live Mammalian Cells by Flow Cytometry Authors:  Claudia Cattoglio, Xavier Darzacq, Robert Tjian and Anders Sejr Hansen, date: 02/20/2020, view: 1485, Q&A: 0
Accurate abundance measurements of cellular proteins are required to achieve a quantitative and predictive understanding of any biological process inside the cell. Existing methods to determine absolute protein abundances are labor-intensive and/or require sophisticated experimental and computational infrastructure (e.g., fluorescence ...
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Preparation of Single Epithelial Cells Suspension from Mouse Mammary Glands Authors:  Heng Sun, Xiaoling Xu and Chuxia Deng, date: 02/20/2020, view: 947, Q&A: 0
Single cell RNA sequencing is a very powerful means for cellular heterogeneous studies and so becoming wildly utilized nowadays. To guarantee the success of such analysis, it is very important, though sometimes difficult, to obtain single cells suspension with high quality, especially from the primary solid organs like mammary glands. Digestion of ...
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Developmental Biology

RNA Sequencing of Single Myofibers from Mus musculus Authors:  Darren M. Blackburn, Felicia Lazure and Vahab D. Soleimani, date: 02/20/2020, view: 1624, Q&A: 0
Whole transcriptome analysis is a key method in biology that allows researchers to determine the effect a condition has on gene regulation. One difficulty in RNA sequencing of muscle is that traditional methods are performed on the whole muscle, but this captures non-myogenic cells that are part of the muscle. In order to analyze only the ...
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Immunology

Skin Transplantation and Lymphoid Organ Analysis in Mice Authors:  Julie Ruer-Laventie, Rajesh Jayachandran, Mathias Schmaler, Simona W. Rossi and Jean Pieters, date: 02/20/2020, view: 932, Q&A: 0
Skin transplantation in mice is an important procedure to evaluate immune responses generated against heterologous grafts, especially given its highly immunogenic nature. In fact, skin is one of the most challenging organs in terms of allograft retention. In this protocol, we provide a detailed procedure for skin grafting using the tail skin as ...
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Molecular Biology

Surface Plasmon Resonance Analysis of the Protein-protein Binding Specificity Using Autolab ESPIRIT Authors:  Pragyan Parimita Rath, Gaurav Anand and Shalini Agarwal, date: 02/20/2020, view: 1080, Q&A: 0
Direct protein-protein interactions are known to regulate a wide range of cellular activities. To understand these contacts one can employ various experimental methods like Dynamic Light Scattering (DLS), Fluorescence Resonance Energy Transfer (FRET), Isothermal titration calorimetry (ITC), Chemical crosslinking, Co-immunoprecipitation (Co-IP), ...
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Super-resolution Microscopy-based Bimolecular Fluorescence Complementation to Study Protein Complex Assembly and Co-localization Authors:  Jingjing Chen, Zulin Yu, Jay R. Unruh, Brian D. Slaughter and Sue L. Jaspersen, date: 02/20/2020, view: 1262, Q&A: 0
Numerous experimental approaches exist to study interactions between two subunits of a large macromolecular complex. However, most methods do not provide spatial and temporal information about binding, which are critical for dissecting the mechanism of assembly of nanosized complexes in vivo. While recent advances in super-resolution ...
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Assessing Self-interaction of Mammalian Nuclear Proteins by Co-immunoprecipitation Authors:  Claudia Cattoglio, Iryna Pustova, Xavier Darzacq, Robert Tjian and Anders S. Hansen, date: 02/20/2020, view: 1211, Q&A: 0
Protein-protein interactions constitute the molecular foundations of virtually all biological processes. Co-immunoprecipitation (CoIP) experiments are probably the most widely used method to probe both heterotypic and homotypic protein-protein interactions. Recent advances in super-resolution microscopy have revealed that several nuclear proteins ...
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Neuroscience

Hippocampal Unicellular Recordings and Hippocampal-dependent Innate Behaviors in an Adolescent Mouse Model of Alzheimer’s disease Authors:  Siddhartha Mondragón-Rodríguez, Benito Ordaz, Erika Orta-Salazar, Sofia Díaz-Cintra, Fernando Peña-Ortega and George Perry, date: 02/20/2020, view: 787, Q&A: 0
Transgenic mice have been used to make valuable contributions to the field of neuroscience and model neurological diseases. The simultaneous functional analysis of hippocampal cell activity combined with hippocampal dependent innate task evaluations provides a reliable experimental approach to detect fine changes during early phases of ...
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Method for Primary Epithelial Cell Culture from the Rat Choroid Plexus Authors:  Valeria Lallai, Amina Ahmed and Christie D. Fowler, date: 02/20/2020, view: 1001, Q&A: 0
The choroid plexus consists of a network of secretory epithelial cells localized throughout the lateral, third and fourth ventricles of the brain. Cerebrospinal fluid (CSF) is generated by the choroid plexus and released into the ventricular environment. This biofluid contains an enriched source of proteins, ions, and other signaling molecules for ...
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Induction of Temporal Lobe Epilepsy in Mice with Pilocarpine Authors:  Muhammad Nauman Arshad and Janice R Naegele, date: 02/20/2020, view: 936, Q&A: 0
In the pilocarpine model of temporal lobe epilepsy (TLE) in rodents, systemic injections of pilocarpine induce continuous, prolonged limbic seizures, a condition termed “Status Epilepticus” (SE). With appropriate doses, many inbred strains of mice show behavioral seizures within an hour after pilocarpine is injected. With the behavioral scoring ...
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Plant Science

Construction of Antisense RNA-mediated Gene Knock-down Strains in the Cyanobacterium Anabaena sp. PCC 7120 Authors:  Amit Srivastava, Anand Ballal, Karl Forchhammer and Anil Kumar Tripathi, date: 02/20/2020, view: 940, Q&A: 0
Anabaena sp. PCC 7120 (hereafter Anabaena) is a model cyanobacterium to study nitrogen fixation, cellular differentiation and several other key biological functions that are analogous in plants. As with any other organism, many genes in Anabaena encode an essential life function and hence cannot be deleted, causing a ...
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Stem Cell

Mesenchymal Stromal Cells Derived from Bone Marrow and Adipose Tissue: Isolation, Culture, Characterization and Differentiation Authors:  Gileade P. Freitas, Alann T. P. Souza, Helena B. Lopes, Rayana L. B. Trevisan, Fabiola S. Oliveira, Roger R. Fernandes, Fernanda U. Ferreira, Felipe A. Ros, Marcio M. Beloti and Adalberto L. Rosa, date: 02/20/2020, view: 1039, Q&A: 0
Since their discovery, mesenchymal stromal cells (MSCs) have received a lot of attention, mainly due to their self-renewal potential and multilineage differentiation capacity. For these reasons, MSCs are a useful tool in cell biology and regenerative medicine. In this article, we describe protocols to isolate MSCs from bone marrow (BM-MSCs) and ...
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