Biotin Pulldown Selection
Biotin Pulldown Selection - Related Videos
11:00
Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
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Cited by 31 •
2018
This report describes a fast and reliable method for validating mRNA targets of cellular miRNAs. The method uses synthetic biotinylated Locked Nucleic Acid (LNA)-based miRNA mimics to capture target mRNA. Subsequently, streptavidin-coated magnetic beads are employed to pulldown the target mRNA for quantification by qPCR polymerase chain reaction.
05:16
Assessing the Internalization of Target Surface Proteins Using a Biotin Derivative
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2025
This video demonstrates a method to assess target protein internalization in the mouse cortical astrocytes using biotinylation, followed by cell lysis, streptavidin-based protein extraction, denaturation, and Western blot analysis to confirm successful internalization.
03:12
An Avidin-Biotin Conjugation Technique for Presenting Target Antigens on Mycobacterium bovis BCG
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2025
The video demonstrates a technique for loading antigens on Mycobacterium bovis BCG to improve its immunogenic properties. The method uses the avidin-biotin system to coat the bacterial surface with exogenous antigens.
04:10
Helicase Activity Measurement of a Target Protein Using Biotin-Labeled RNA Duplexes
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2025
In this video, we demonstrate the procedure to determine the helicase activity of a target protein to unwind the biotin-labeled dsRNA substrate. The activity of the enzyme was identified by analyzing the electrophoretic mobility shift, followed by a chemiluminescence assay using chemiluminescent enzyme-conjugated streptavidin.
16:33
Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange (ABE)
0 Views •
Cited by 108 •
2013
The reversible addition of palmitate to proteins is an important regulator of intracellular protein trafficking. This is of particular interest in neurons where many synaptic proteins are palmitoylated. We utilize a simple biochemical method to detect palmitoylated proteins in cultured neurons, which can be adapted for multiple cell types and tissues.
FAQs
Selectivity comes from attaching biotin to the nucleic acid, protein, or molecular complex of interest and using streptavidin-coated beads as the capture surface. The tagged material is retained through this affinity interaction, whereas other mixture components can remain unbound. This enrichment creates a concentrated starting material for examining genetic interactions.
Washing separates material that did not remain associated with the bead-bound capture system from the targets and associated molecules retained for analysis. In a genetic pulldown, this step helps distinguish selected DNA or RNA, or proteins linked to regulatory regions, from the original mixture. The retained fraction provides the material used in downstream assays.
Biotin Pulldown Selection can be adapted to enrich selected DNA sequences, RNA sequences, proteins that bind regulatory regions, or larger molecular complexes. Because the captured material may represent either a target or its associated partners, the approach supports questions about nucleic-acid enrichment and genetic interactions. This flexibility makes it relevant across genetics experiments.
A typical workflow begins by tagging the selected target with biotin, introducing it to streptavidin-coated beads, and separating bead-associated material from the mixture. Washing removes unbound components, after which the retained targets or associated molecules are recovered for electrophoresis, sequencing, or mass spectrometry. The chosen assay determines what is measured.
It is particularly useful when a selected genetic molecule or interaction is difficult to detect directly in a complex mixture. By enriching low-abundance targets, the approach can make downstream examination more informative. In genetics, that benefit supports studies of gene regulation, genome organization, and molecular interactions involving selected DNA or RNA sequences.
Downstream analysis can show what was enriched after capture and washing. Electrophoresis provides a way to examine the retained material, whereas sequencing can characterize selected nucleic acids and mass spectrometry can identify associated proteins. These readouts connect the physical pulldown to questions about gene regulation, genome organization, or molecular interactions.