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Protocol
Co-immunoprecipitation for Assessing Protein–Protein Interactions in Agrobacterium-Mediated Transient Expression System in Nicotiana benthamiana
The characterization of protein–protein interactions (PPI) often provides functional information about a target protein. Yeast-two-hybrid (Y2H) and luminescence/fluorescence-based detections, therefore, have b...
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Protocol
Compact Eight-Channel Light-Sensing System for Bioassays
The present protocol introduces a new instrumental setup as a luminometer to simultaneously measure eight light samples with high sensitivity. The system consists of 8-channel photomultiplier tubes (8-PMTs) wi...
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Protocol
Molecular Tension Probe for In Vitro Bioassays
Cell-free bioassays (CFBs) provide their own distinctive merits over cell-based bioassays (CBBs) including (i) rapid and on-site applicability, (ii) long-term utility, and (iii) bioanalytical versatility. The ...
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Protocol
One-Channel Microsliding Luminometer for Quantifying Low-Energy Bioluminescent Lights
A high-throughput quantitative determination of multiple light-emitting samples is a virtue of many light determination systems. In this chapter, we introduce a compact and efficient light determination system...
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Protocol
Quantitative Imaging of Retinoic Acid Activities in Living Mammalian Cells
Retinoic acid (RA) is an intriguing metabolite that is necessary for embryonic development and differentiation in vertebrates. The present protocol demonstrates how to image RA activities indirectly in mammali...
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Protocol
Visible Light Bioluminescence Imaging Platform for Animal Cell Imaging
The present protocol introduces a visible light bioluminescence imaging (BLI) platform together with 12 novel coelenterazine (CTZ) analogues and luciferase sets. We exemplify to create diverse hues of biolumin...
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Protocol
Near-Infrared Bioluminescence Imaging of Animal Cells with Through-Bond Energy Transfer Cassette
is the most general substrate for marine . The present protocol introduces a near-infrared (NIR ) bioluminescence (BL) of mammalian cells with a cyanine-5 (Cy5) dye-conjugated CTZ . This unique Cy5-conjuga...
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Protocol
Highly Bright NIR-BRET System for Imaging Molecular Events in Live Cells
The present protocol demonstrates a novel mammalian cell imaging platform exerting a bioluminescence resonance energy transfer (BRET) system. This platform achieves a ~300 nm blue-to-near infrared shift of the...
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Protocol
Ligand-Activatable BRET9 Probes for Imaging Molecular Events in Living Mammalian Cells
Bioluminescence resonance energy transfer (BRET) is a commonly used assay system for studying protein-protein interactions. The present protocol introduces a conceptually unique ligand-activatable BRET system ...
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Protocol
A New Lineage of Artificial Luciferases for Mammalian Cell Imaging
The present protocol introduces a new lineage of artificial luciferases (ALucs) with unique optical properties for mammalian cell imaging. The primary candidate sequence was first created with a sequence logo ...
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Protocol
Azide- and Dye-Conjugated Coelenterazine Analogues for Imaging Mammalian Cells
Coelenterazine (CTZ) is a common substrate to most marine luciferases and photoproteins. The present protocol introduces mammalian cell imaging with nine novel dye- and azide-conjugated CTZ analogues, which we...
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Protocol
Luciferase-Specific Coelenterazine Analogues for Optical Cross Talk-Free Bioassays
Spectral overlaps in fluorescence (FL) and bioluminescence (BL) commonly cause optical cross talks. The present protocol introduces five different lineages of coelenterazine (CTZ) analogues, which have selecti...
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Protocol
High-Throughput Targeted Transcriptional Profiling of Defense Genes Using RNA-Mediated Oligonucleotide Annealing, Selection, and Ligation with Next-Generation Sequencing in Arabidopsis
Tracking RNA transcription has been one of the most powerful tools to gain insight into the biological process. While a wide range of molecular methods such as northern blotting, RNA-seq, and quantitative RT-P...
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Protocol
Methods for the Study of Entotic Cell Death
Entosis is a mechanism of cell competition occurring in cancers that involves the engulfment and killing of neighboring cells. The death of ingested cells, called entotic cell death, usually occurs in a non-ap...
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Protocol
Identification of Long Noncoding RNAs in the Develo** Endosperm of Maize
Maize endosperm consists of three distinct types of tissues, including the starchy endosperm (SE), the basal endosperm transfer cell layer (BETL), and the aleurone cell layer (AL). Compartmentalization of thes...
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Protocol
A Bioluminescence Assay System for Imaging Metal Cationic Activities in Urban Aerosols
A bioluminescence-based assay system was fabricated for an efficient determination of the activities of air pollutants. The following four components were integrated into this assay system: (1) an 8-channel as...
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Protocol
A Multichannel Bioluminescence Determination Platform for Bioassays
The present protocol introduces a multichannel bioluminescence determination platform allowing a high sample throughput determination of weak bioluminescence with reduced standard deviations. The platform is d...
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Protocol
Single-Chain Probes for Illuminating Androgenicity of Chemicals
The present protocol introduces a single-chain probe carrying a functional peptide in the N-terminal domain of the androgen receptor (AR NTD) for illuminating androgenicity of ligands. In the single-chain prob...
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Protocol
Circular Permutation Probes for Illuminating Phosphorylation of Estrogen Receptor
The present protocol demonstrates a new strategy for imaging ligand-triggered protein phosphorylation using circularly permutated luciferases (cpLuc): (1) a luciferase is first fragmented into two segments for...
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Protocol
Multicolor Imaging of Bifacial Activities of Estrogens
The present protocol introduces multicolor imaging of bifacial activities of an estrogen. For the multicolor imaging, the authors fabricated two single-chain probes emitting green or red bioluminescence (named...