63 Result(s)
within
Protocol
Yu** Lai
Life Sciences
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Protocol
Protein–Protein Interactions: Co-immunoprecipitation
Proteins often do not function as single substances but rather as team players in a dynamic network. Growing evidences show that protein–protein interactions are crucial in many biological processes in living ...
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Protein–Protein Interactions: Yeast Two Hybrid
The yeast is a powerful and commonly used genetic tool to investigate the interaction between artificial fusion proteins inside the nucleus of yeast. Here, we describe how to use the Matchmaker GAL4-based ...
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Quantification of C. neoformans Capsule Diameter
The polysaccharide capsule of Cryptococcus neoformans is the primary virulence factor and one of the most commonly studied aspects of this pathogenic yeast. Capsule size varies widely between strains, has the abi...
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Dissecting the Organization of a β-Barrel Assembly Machinery (BAM) Complex by Neutron Reflectometry
The β-barrel assembly machinery (BAM) complex in Gram-negative bacteria facilitates the assembly of β-barrel proteins into the outer membrane. Understanding the protein-protein interactions within this complex...
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RNA-Binding Protein-Mediated mRNA Deadenylation in Mammalian Cell Extracts
Removal of the poly(A) tail, or deadenylation, is a crucial step in destabilizing mRNAs in eukaryotes. In this chapter, we describe a cell-free deadenylation assay that uses cytoplasmic cell extracts from huma...
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In Situ Structure Determination of Bacterial Surface Nanomachines Using Cryo-Electron Tomography
Bacterial surface nanomachines are often refractory to structural determination in their intact form due to their extensive association with the cell envelope preventing them from being properly purified for t...
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Single-Molecule Imaging of the Phase Separation-Modulated DNA Compaction to Study Transcriptional Repression
Liquid-liquid phase separation (LLPS) often induces the formation of biomolecule condensates at the cellular level. The importance of this phenomenon has been demonstrated in many important biological function...
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Antibody Phage Display
The application of antibodies has transcended across many areas of work but mainly as a research tool, for diagnostic and for therapeutic applications. Antibodies are immunoproteins from vertebrates that have ...
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Cell Biological Analyses of Anther Morphogenesis and Pollen Viability in Arabidopsis and Rice
Major advances have been made in our understanding of anther developmental processes in flowering plants through a combination of genetic studies, cell biological technologies, biochemical analyses, microarray...
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Flower Development in Arabidopsis
Like in other angiosperms, the development of flowers in Arabidopsis starts right after the floral transition, when the shoot apical meristem (SAM) stops producing leaves and makes flowers instead. On the flan...
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Construction of Naïve and Immune Human Fab Phage Display Library
Phage display has been applied successfully for the rapid isolation of monoclonal antibodies against various targets including infectious diseases, autoantigens, cancer markers, and even small molecules. The m...
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Streptavidin-Coated Solid-Phase Extraction (SPE) Tips for Antibody Phage Display Biopanning
Phage display is a technique that allows the presentation of unique proteins on the surface of bacteriophages. The phage particles are usually screened via repetitive rounds of antigen-guided selection and pha...
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Identification of Plant Transcription Factor DNA-Binding Sites Using seq-DAP-seq
The identification of genome-wide transcription factor binding sites (TFBS) is a critical step in deciphering gene and transcriptional regulatory networks. However, determining the genome-wide binding of speci...
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Targeted Mass Spectrometry-Based Proteomics Method to Quantify Placental Extracellular Vesicles
Extracellular vesicles (EVs) carry a wide range of molecules, such as proteins, RNAs, and DNA. EVs are secreted from a wide range of cells, including placental cells. Interestingly, EVs secreted from placental...
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AGROBEST: A Highly Efficient Agrobacterium-Mediated Transient Expression System in Arabidopsis Seedlings
Agrobacterium-mediated transient transformation for gene expression is a simple and fast method to analyze transgene functions in plants. Agroinfiltration in leaves of Nicotiana benthamiana is a common method for...
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Modulation of VEGFA Signaling During Heart Regeneration in Zebrafish
Over the last decades, myocardial infarction and heart failure have accounted every year for millions of deaths worldwide. After a coronary occlusion, the lack of blood supply to downstream muscle leads to cel...
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Probing Transcriptome-Wide RNA Structural Changes Dependent on the DEAD-box Helicase Dbp2
RNA helicases function in all aspects of RNA biology mainly through remodeling structures of RNA and RNA-protein (RNP) complexes. Among them, DEAD-box proteins form the largest family in eukaryotes and have be...
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Knocking Out the Wall: Revised Protocols for Gene Targeting in Physcomitrella patens
The moss Physcomitrella patens has become established as a model for investigating plant gene function due to the feasibility of gene targeting. The chemical composition of the P. patens cell wall is similar to t...
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Isolation of Synthetic Antibodies Against BCL-2-Associated X Protein (BAX)
The BCL-2 protein family plays central roles in the mitochondrial pathway of cell apoptosis. The BCL-2-Associated X protein (BAX), along with other proapoptotic proteins, induces cell death in response to a va...
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Metaproteomics Study of the Gut Microbiome
Proteomics is a widely used method for defining the protein composition of a complex sample. As this approach allows for identification and quantification of proteins across a broad dynamic range as well as de...
