63 Result(s)
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Protocol
Large-Scale Purification of Natural Ice-Binding Proteins by the Falling Water Ice Purification Method
We describe here a generic technique for purification of ice-binding proteins (IBPs) using a commercial ice machine. The method, which is called falling water ice purification (FWIP) [1], is based on the affinity...
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Protocol
Structure-Guided and Phage-Assisted Evolution of Therapeutic Antibodies to Reverse On-Target Point Mutation-Mediated Resistance
Resistance to therapeutic antibodies caused by on-target point mutations is a major obstacle in anticancer therapy, creating an “unmet clinical need.” To tackle this problem, researchers are develo** new gen...
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Protocol
Preparation of Bioconjugates of Chimeric M13 Phage and Gold Nanorods
Phage-nanomaterial conjugates are functional bio-nanofibers with various applications. While phage display can select for phages with desired genetically encoded functions and properties, nanomaterials can end...
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Protocol
Enzymatic Preparation of DNA with an Expanded Genetic Alphabet Using Terminal Deoxynucleotidyl Transferase and Its Applications
Efficient preparation of DNA oligonucleotides containing unnatural nucleobases (UBs) that can pair with their cognates to form unnatural base pairs (UBPs) is an essential prerequisite for the application of UB...
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Protocol
Unnatural Amino Acid Engineering for Intracellular Delivery of Protein Therapeutics
Protein drugs are a critically important therapeutic modality due to the sophisticated binding recognition, catalytic properties, and disease relevance of proteins. There is a clear need for new strategies abl...
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Protocol
Engineering E2 Bionanoparticles for Targeted Delivery of Chemotherapeutics to Breast Cancer Cells
Naturally occurring protein nanocages are promising drug carriers as both the interior and exterior can be decorated for drug encapsulation and cell targeting. To provide surface functionalization, we added a ...
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Protocol
Visualization of Engineered M13 Phages Bound to Bacterial Targets by Transmission Electron Microscopy
The filamentous phage M13 is one of the most well-studied and characterized phages, particularly since it was introduced as a scaffold for phage display, a technique to express and evolve fusion proteins on th...
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Protocol
Engineering Hepatitis B Virus (HBV) Protein Particles for Therapeutic Delivery
Nature provides an abundance of proteins whose structures and reactivity have been perfected through evolution to perform specific tasks necessary for biological function. The structural and functional propert...
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Protocol
Generation and Characterization of In Vitro Transcribed mRNA
Here we describe the in vitro preparation of mRNA from DNA templates, including setting up the transcription reaction, mRNA cap**, and mRNA labeling. We then describe methods used for mRNA characterization, ...
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Protocol
Construction of Xylose-Utilizing Cyanobacterial Chassis for Bioproduction Under Photomixotrophic Conditions
Xylose is a major component of lignocellulose and the second most abundant sugar present in nature after glucose; it, therefore, has been considered to be a promising renewable resource for the production of b...
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Protocol
Genetically Encoded Noncanonical Amino Acids in Proteins to Investigate Lysine Benzoylation
Posttranslational modifications (PTMs) of lysine residues are major regulators of gene expression, protein–protein interactions, and protein localization and degradation. Histone lysine benzoylation is a recen...
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Protocol
Preparation and Fractionation of Heterogeneous Aβ42 Oligomers with Different Aggregation Properties
Deposition of amyloid-β (Aβ) aggregates in the form of amyloid plaques is a central feature of Alzheimer’s disease. The end products of Aβ aggregation are amyloid fibrils. Soluble Aβ aggregates called oligomer...
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Protocol
Single-Cell Micropatterning by Non-fouling Hydrogels
Micropatterned substrate is a unique method for studying cell biology at the single-cell level. Using photolithography to create binary patterns of cell-adherent peptide surrounding by non-fouling cell-repelle...
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Protocol
Genetic Code Expansion in Mammalian Cells Through Quadruplet Codon Decoding
Genetic code expansion enables the site-specific incorporation of noncanonical amino acids (ncAAs) into proteins both in vitro and in vivo. In addition to a widely applied nonsense suppression strategy, the us...
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Protocol
Site-Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cells
Protein (PTS) plays a crucial role in numerous extracellular protein-protein interactions. It is involved in diverse physiological processes and the development of human diseases, including AIDS and cancer. ...
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Protocol
Cross-Seeding Assay in the Investigation of the Amyloid Core of Prion Fibrils
Amyloidogenesis, self-propagation of protein or peptide monomers to amyloid fibrils, has been linked to incurable pathogenesis of neurodegenerative diseases such as Alzheimer’s disease and prion diseases. Inve...
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Protocol
Cross-Linking Mass Spectrometry Analysis of Metastable Compact Structures in Intrinsically Disordered Proteins
Protein assembly into beta-sheet-rich amyloids is a common phenomenon in neurodegenerative diseases including Alzheimer’s (AD) and Parkinson’s (PD). The proteins implicated in amyloid deposition are often intr...
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Protocol
Selective Inhibition of Kinase Activity in Mammalian Cells by Bioorthogonal Ligand Tethering
Enzymes are critical for cellular functions, and malfunction of enzymes is closely related to many human diseases. Inhibition studies can help in deciphering the physiological roles of enzymes and guide conven...
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Protocol
Network-Based Structural Alignment of RNA Sequences Using TOPAS
TOPAS (TOPological network-based Alignment of Structural RNAs) is a network-based alignment algorithm that predicts structurally sound pairwise alignment of RNAs. In order to take advantage of recent advances in ...
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Protocol
Microfluidics-Enabled Isolation and Single-Cell Analysis of Circulating Tumor Cells
Microfluidic platforms enable the enrichment and analysis of circulating tumor cells (CTCs), a potential biomarker for cancer diagnosis, prognosis, and theragnosis. Combined with immunocytochemistry/immunofluo...
