Biological Engineering

Categories

    Protocols in Current Issue
    Purification of Crimean Congo Hemorrhagic Fever Virus (CCHFV) Nucleocapsid Protein Using Detergent Gradient and Free Thawing
    Authors:  Austin Royster and Sheema Mir, date: 08/05/2022, view: 141, Q&A: 0
    [Abstract]

    Protein aggregation remains a major challenge in the purification of recombinant proteins in both eukaryotic and prokaryotic expression systems. One such protein is the nucleocapsid protein of Crimean Congo Hemorrhagic fever virus (CCHFV), which has high aggregation tendency and rapidly precipitates upon purification by NiNTA chromatography. Using

    ...
    Isolation and ex vivo Expansion of Limbal Mesenchymal Stromal Cells
    Authors:  Naresh Polisetti, Lyne Sharaf, Thomas Reinhard and Günther Schlunck, date: 07/20/2022, view: 478, Q&A: 0
    [Abstract]

    Limbal mesenchymal stromal cells (LMSC), a cellular component of the limbal stem cell niche, have the capability of determining the fate of limbal epithelial progenitor cells (LEPC), which are responsible for the homeostasis of corneal epithelium. However, the isolation of these LMSC has proven to be difficult due to the small fraction of LMSC in

    ...
    VirScan: High-throughput Profiling of Antiviral Antibody Epitopes
    Authors:  Ellen L. Shrock, Christine L. Shrock and Stephen J. Elledge, date: 07/05/2022, view: 1090, Q&A: 0
    [Abstract]

    Profiling the specificities of antibodies can reveal a wealth of information about humoral immune responses and the antigens they target. Here, we present a protocol for VirScan, an application of the phage immunoprecipitation sequencing (PhIP-Seq) method for profiling the specificities of human antiviral antibodies. Accompanying this protocol is

    ...
    A Robust Nanoparticle-based Magnetic Separation Method for Intact Lysosomes
    Authors:  The Son Le, Mari Takahashi and Shinya Maenosono, date: 07/05/2022, view: 1258, Q&A: 0
    [Abstract]

    Lysosome isolation is a preresiquite for identifying lysosomal protein composition by mass spectroscopic analysis, to reveal lysosome functions, and their involvement in some diseases. Magnetic nanoparticle-based fractionation has received great attention for lysosome isolation, owing to its high efficiency, purity, and preservation of lysosomal

    ...
    Serological Measurement of Poly-IgA Immune Complex Levels in IgA Nephropathy and IgA Vasculitis
    Authors:  Xue Zhang, Jicheng Lv, Pan Liu, Xinfang Xie, Xinyan Li, Hong Zhang and Jing Jin, date: 07/05/2022, view: 1159, Q&A: 0
    [Abstract]

    Both IgA nephropathy and IgA vasculitis, formerly known as Henoch-Schӧnlein purpura, are immune deposition diseases. IgA nephropathy is caused by the deposition of aberrantly formed poly-IgA complexes from blood circulation to the kidney glomerulus; IgA vasculitis is characterized by IgA-dominant immune deposits to small vessels of the skin

    ...
    In vivo Characterization of Endogenous Protein Interactomes in Drosophila Larval Brain, Using a CRISPR/Cas9-based Strategy and BioID-based Proximity Labeling
    Authors:  Ezgi Uçkun, Georg Wolfstetter, Johannes Fuchs and Ruth H. Palmer, date: 07/05/2022, view: 1142, Q&A: 0
    [Abstract]

    Understanding protein-protein interactions (PPIs) and interactome networks is essential to reveal molecular mechanisms mediating various cellular processes. The most common method to study PPIs in vivo is affinity purification combined with mass spectrometry (AP–MS). Although AP–MS is a powerful method, loss of weak and transient

    ...
    Protocol for High Throughput Screening of Antibody Phage Libraries
    [Abstract]

    Phage display is a proven and widely used technology for selecting specific antibodies against desired targets. However, an immense amount of effort is required to identify and screen the desired positive clones from large and diverse combinatorial libraries. On the other hand, the selection of positive binding clones

    ...
    Preparation of Cas9 Ribonucleoproteins for Genome Editing
    Authors:  Sheng-Wei Lin, Viet Quoc Nguyen and Steven Lin, date: 05/20/2022, view: 1652, Q&A: 0
    [Abstract]

    Genome editing by the delivery of pre-assembled Cas9 ribonucleoproteins (Cas9 RNP) is an increasingly popular approach for cell types that are difficult to manipulate genetically by the conventional plasmid and viral methods. Cas9 RNP editing is robust, precise, capable of multiplexing, and free of genetic materials. Its transient presence in

    ...
    Immunomagnetic Isolation and Enrichment of Microvascular Endothelial Cells from Human Adipose Tissue
    [Abstract]

    Human adipose tissue-resident microvascular endothelial cells are not only garnering attention for their emergent role in the pathogenesis of obesity-related metabolic disorders, but are also of considerable interest for vascular tissue engineering due, in part, to the abundant, accessible, and uniquely dispensable nature of the tissue. Here, we

    ...
    Activation of Mitochondrial Ca2+ Oscillation and Mitophagy Induction by Femtosecond Laser Photostimulation
    Authors:  Xiaoying Tian and Hao He, date: 04/05/2022, view: 566, Q&A: 0
    [Abstract]

    Ultra-precise stimulation solely to individual mitochondria, without any influence to the whole cell, is quite difficult by traditional biochemical reagents. In mitophagy research, the mitochondria and even the whole cell usually suffer irreversible and great damage caused by treatment with potent chemicals. In this protocol, we present the

    ...



    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.