79 Result(s)
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Protocol
Use of Reductive Amination to Produce Capsular Polysaccharide-Based Glycoconjugates
Reductive amination is a relatively simple and convenient strategy for coupling purified polysaccharides to carrier proteins. Following their synthesis, glycoconjugates can be used to assess the protective cap...
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Protocol
Examining Phosphatases Through Immunofluorescent Microscopy
Immunofluorescent microscopy enables the examination of cellular expression and localization of proteins. Cellular localization can often impact protein function, as certain molecular interactions occur in spe...
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Protocol
Application of the CRISPR/Cas9 Gene Editing Method for Modulating Antibody Fucosylation in CHO Cells
Genetic engineering plays an essential role in the development of cell lines for biopharmaceutical manufacturing. Advanced gene editing tools can improve both the productivity of recombinant cell lines as well...
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Protocol
Molecular Docking of Intrinsically Disordered Proteins: Challenges and Strategies
Intrinsically disordered proteins (IDPs) are a novel class of proteins that have established a significant importance and attention within a very short period of time. These proteins are essentially characteri...
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Protocol
Characterization of Crystals Isolated from the Caesalpinia sappan Seeds: A Comparative In Silico Analysis on Bioactivity of Sappan Diterpenoids
Objective. Caesalpinia sappan is a genus of plants belonging to the family Fabaceae found in East and Southeast Asia. Pharmacologically, species of this sort display anti-inflammatory, cytotoxic, antibacterial, a...
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Protocol
A SINC-Seq Protocol for the Analysis of Subcellular Gene Expression in Single Cells
Microfluidic devices offer precise control of single cells and molecules by liquid flows, downsizing tools to allow us to perform single-cell assays at unprecedented resolutions and minimizing contamination. I...
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Protocol
Inflammasome Activation in Human Macrophages: IL-1β Cleavage Detection by Fully Automated Capillary-Based Immunoassay
Interleukin (IL)-1β is a key mediator of inflammation and activates via pattern recognition receptors (PRR) of the inflammasome family by proteolytic maturation. Proteolysis is driven by proteases such as casp...
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Protocol
Production of α-Synuclein Fibrillar-Specific scFv from Inclusion Bodies
Recombinant antibody fragments such as Fab, scFvs, and diabodies against α-syn have become a viable alternative to the conventional full-length antibodies in immunotherapeutic approaches due to their benefits ...
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Protocol
Monitoring Kinetics of pH-Dependent Aggregation and Disaggregation of the Pmel17 Repeat Domain
The premelanosomal protein (Pmel17) is a human functional amyloid that promotes pigmentation by serving as a scaffold for melanin polymerization. This occurs within the melanosome, where Pmel17 is first proteo...
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Protocol
Flow Cytometry-Based Detection of Siglec Ligands
Siglecs are a family of immunomodulatory cell surface receptors present on white blood cells. Binding to cell surface sialic acid-containing glycans modulates the proximity of Siglecs to other receptors that t...
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Protocol
Cryo-electron Microscopy of Protein Cages
Protein cages are one of the most widely studied objects in the field of cryogenic electron microscopy—encompassing natural and synthetic constructs, from enzymes assisting protein folding such as chaperonin t...
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Protocol
Analysis of CCN4/WISP1 Effects on Joint Tissues Using Gain- and Loss-of-Function Approaches
The matricellular protein Wnt-induced secreted protein 1 (WISP1 ) is the fourth member of the CCN family of proteins, which has been shown to affect tissues of the musculoskeletal system. In the context of the...
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Protocol
Directed Evolution of Fluorescent Proteins in Bacteria
Directed evolution has revolutionized the way scientists create new biomolecules not found in nature. Error-prone polymerase chain reaction (PCR) introduces random mutations and was used to evolve jellyfish an...
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Protocol
Using FRET-Based Biosensor Cells to Study the Seeding Activity of Tau and α-Synuclein
Two fluorescence resonance energy transfer (FRET)-based biosensor cell lines developed several years ago by the Diamond group (University of Texas, Southwestern) have allowed convenient, sensitive, and specifi...
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Protocol
In Vitro Models on Inhibition of Inflammatory Mediators by Ethanolic Extract of Careya arborea Roxb. Bark in Lipopolysaccharide-Induced RAW 264.7 Cell Lines
Inflammation is a normal protective response to tissue injury caused by physical trauma, noxious chemicals, or microbiological agents. When this normal physiological process becomes deregulated, it can become ...
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Protocol
Qualitative and Quantitative Characterization of Protein–Carbohydrate Interactions by NMR Spectroscopy
Solution-state nuclear magnetic resonance (NMR) spectroscopy can be used to monitor protein–carbohydrate interactions. Two-dimensional 1H-15N heteronuclear single quantum coherence (HSQC)-based techniques describ...
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Protocol
Semi-Automated Lectin Magnetic Bead Array (LeMBA) for Translational Serum Glycoprotein Biomarker Discovery and Validation
Aberrant protein glycosylation is a characteristic of diverse diseases which has been explored as biomarkers. To support translational serum glycoprotein biomarker discovery and validation, we developed a semi...
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Protocol
Map** Phase Diagram of Tau-RNA LLPS Under Live Cell Coculturing Conditions
Protein liquid-liquid phase separation (LLPS) has been associated with biological functions and pathological aggregation. Map** the phase separation conditions is the first step to identify and quantify the ...
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Protocol
Combination of Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and Substrate Trap** for the Detection of Transient Protein Interactions
Antibody-based affinity purification is a recognized method for use in studying protein–protein interactions. There are four different classes of proteins that are typically identified with such affinity purif...
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Protocol
Mass Spectrometry-Based Untargeted Metabolomics and Lipidomics Platforms to Analyze Cell Culture Extracts
Metabolites represent the most downstream level of the cellular organization. Hence, an in vitro untargeted metabolomics approach is extremely valuable to deepen the understanding of how endogenous metabolites...
