Biotinylation Reagent
Biotinylation Reagent - Related Videos
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...
10:17
Preparing Anhydrous Reagents and Equipment
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2023
Source: Laboratory of Dr. Dana Lashley - College of William and Mary Demonstrated by: Timothy Beck and Lucas Arney Many reactions in organic chemistry are moisture-sensitive and must be carried out under careful exclusion of water. In these cases the reagents have a high affinity to react with water from the atmosphere and if left exposed the desired reaction will not take place or give poor yields, because the reactants are chemically altered. In order to prevent undesired reactions with H2O...
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.
11:32
Measuring Plasma Membrane Protein Endocytic Rates by Reversible Biotinylation
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Cited by 23 •
2009
Regulated endocytosis governs the cell surface expression levels of the majority of membrane proteins. Here we utilize reducible, membrane impermeant biotinylation reagents to measure the endocytic rate of the dopamine transporter (DAT), a polytopic membrane protein. The method facilitates a straightforward approach to measuring the endocytic rate of most plasma membrane proteins.
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.
FAQs
The reactive group determines which biomolecular site is labeled. NHS esters target lysine residues and N-termini, whereas maleimides react with accessible cysteine thiols under suitable conditions. This distinction allows researchers to choose a reagent according to the available chemical groups on a protein, antibody, peptide, or other biomolecule, helping tailor labeling to the intended assay or purification use.
After labeling, biotin provides a recognition site for avidin or streptavidin, which binds it with high affinity. That interaction connects the biomolecule to detection, capture, or purification formats without requiring the original protein or antibody to carry those functions itself. Consequently, the same labeling strategy can support signal generation, affinity-based isolation, or analytical enrichment.
Labeling conditions must be controlled because the modification should add a useful biotin handle without compromising the biomolecule’s activity. Preserving antibody or antigen function is important for specific recognition in downstream assays. Consistent conditions also improve assay sensitivity and reproducibility, making results more comparable across experiments and reducing variability attributable to the labeling step.
A typical workflow begins by selecting a reagent whose reactive group matches the biomolecule’s available lysine, N-terminal, or cysteine thiol sites. The labeling reaction is then performed under suitable, controlled conditions. The resulting biotinylated molecule can be paired with avidin or streptavidin for detection, capture, purification, or other assay formats supported by the experiment.
Biotinylated antibodies and antigens support several established workflows in immunology and infection research. They can contribute to enzyme-linked assays, flow cytometry, affinity purification, and pathogen detection. The selected biomolecule determines the recognition element, while the attached biotin enables interaction with avidin or streptavidin-based detection or capture systems.
In pathogen detection, a biotinylated antibody or antigen can link molecular recognition to an avidin or streptavidin-based assay system. Antibodies provide a way to target relevant molecular features, while antigens can serve as assay components for immune recognition. This separation of recognition and detection helps adapt the same labeling principle to different infection and immunology formats.