Biotinylation Selection
Biotinylation Selection - Related Videos
10:43
Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes
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2019
Here we present a method to select for novel variants of the E. coli biotin-protein ligase BirA that biotinylates a specific target peptide. The protocol describes the construction of a plasmid for the bacterial display of the target peptide, generation of a BirA library, selection and characterization of BirA variants.
09:13
Cell-surface Biotinylation Assay
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2023
A cell can regulate the amount of particular proteins on its cell membrane through endocytosis, following which cell surface proteins are effectively sequestered in the cytoplasm. Once within a cell, these surface proteins can be either destroyed or “recycled” back to the membrane. The cell surface biotinylation assay provides researchers with a way to study these phenomena. The technique makes use of a derivative of the small molecule biotin, which can label surface proteins and then be...
11:32
Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen
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Cited by 20 •
2017
Here, we detail the method of Sequencing of Psoralen crosslinked, Ligated, and Selected Hybrids (SPLASH), which enables genome-wide mapping of intramolecular and intermolecular RNA-RNA interactions in vivo. SPLASH can be applied to study RNA interactomes of organisms including yeast, bacteria and humans.
06:43
In Vivo Proximity Biotinylation for Protein Interaction Studies in Paramecium tetraurelia
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Cited by 1 •
2025
The protocol presents a method for in vivo covalent attachment of biotin to proteins based on their proximity to a biotin ligase fused to a protein of interest. This modification allows for a selective enrichment of the proteins using streptavidin beads as needed in protein interaction studies.
12:03
Purification of Biotinylated Cell Surface Proteins from Rhipicephalus microplus Epithelial Gut Cells
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Cited by 3 •
2017
A modified density centrifugation gradient-based methodology was utilized to isolate epithelial cells from Rhipicephalus microplus gut tissue. Surface-bound proteins were biotinylated and purified through streptavidin magnetic beads for utilization in downstream applications.
FAQs
The exceptional strength of the biotin-streptavidin interaction allows labeled targets to remain associated with immobilized streptavidin while unbound material is removed. This produces selective enrichment rather than simple detection of biotin. Researchers can then recover captured proteins, peptides, cells, or other targets for downstream analysis of their identity and biomolecular interactions.
Covalent attachment keeps the biotin label linked to the protein, peptide, cell, or other target during the selection process. The labeled material can therefore be presented to immobilized streptavidin for capture. Using this shared labeling principle allows the approach to enrich different classes of biological targets within the same affinity-based framework.
Washing removes material that has not been retained by the immobilized streptavidin, reducing unbound background in the selected fraction. The outcome depends on the contrast between labeled targets that remain captured and material that is removed. This separation supports more focused detection, purification, and characterization of the enriched molecules or cells.
A typical workflow begins by covalently labeling the selected target with biotin. The labeled material is then applied to immobilized streptavidin so the biotin-bearing targets can be captured. Researchers wash away unbound material and retain the captured fraction for analysis. The same sequence can be applied to proteins, peptides, cells, or other targets.
Researchers can use Biotinylation Selection to enrich antigens, antibodies, receptors, and host-pathogen interaction partners. These applications make the method useful when a study requires selective recovery of immune-related molecules or complexes from other material. The enriched targets can support investigations of pathogen recognition, immune targeting, or molecular interactions associated with invasion.
Selected material can support sensitive detection, purification, and characterization of biomolecular interactions. In infection research, analyzing enriched antigens, receptors, or host-pathogen partners may help identify immune targets and clarify how pathogens are recognized or engage host molecules. The method therefore connects affinity-based recovery with mechanistic studies of immunological and infectious processes.