Affinity Enrichment
Affinity Enrichment - Related Videos
12:11
Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
0 Views •
Cited by 9 •
2020
This workflow describes the performance of time- and cost-efficient enrichment of multiple protein post-translational modifications (PTMs) simultaneously for quantitative global proteomic analysis. The protocol utilizes peptide-level PTM enrichment with multiple conjugated antibodies, followed by data-independent acquisition mass spectrometry analysis to gain biological insights into PTM crosstalk.
03:07
Electron Affinity
0 Views •
2020
The electron affinity (EA) is the energy change for adding an electron to a gaseous atom to form an anion (negative ion). This process can be either endothermic or exothermic, depending on the element. Many of these elements have negative values of EA, which means that energy is released when the gaseous atom accepts an electron. However, for some elements, energy is required for the atom to become negatively charged, and the value of their EA is positive. Just as with ionization energy,...
01:41
Affinity and Avidity
0 Views •
2019
Overview Antibodies bind to toxins or substances on the surface of cells, bacteria, viruses, or fungi. The substance is called an antigen, and the precise binding site is the epitope. The strength of the antibody-epitope interaction is called affinity. When an antibody binds an antigen by multiple epitopes, the cumulative strength of the interaction is called avidity. The strength of the interaction influences the elicited immune response. The Adaptive Immune System Increases Efficiency by...
01:34
Environmental Enrichment for Rodents
0 Views •
2026
All animal procedures described here must be conducted in accordance with institutional animal ethics guidelines and approved by IACUC. All procedures must follow the principles of the 3Rs—Replacement, Reduction, and Refinement—and must be performed by trained personnel. Environmental enrichment involves objects and practices that encourage natural behaviors, improve psychological well-being, and promote better welfare in laboratory rodents. Environmental enrichment should be tailored to the...
14:47
Identification of MyoD Interactome Using Tandem Affinity Purification Coupled to Mass Spectrometry
0 Views •
Cited by 5 •
2016
MyoD is a myogenic transcription factor with a strong capacity to induce myogenic transdifferentiation of many fully differentiated non-muscle cell lines. The epigenetic mechanisms involved in this transdifferentiation are largely unknown. Here we describe a double-affinity purification method followed by mass spectrometry to exhaustively characterize MyoD partners.
FAQs
The immobilized ligand serves as the selective capture component of the matrix. Its binding relationship with the target allows that molecule to remain associated with the matrix while other sample constituents do not bind under the chosen conditions. This molecular recognition enables enrichment from a complex biochemical mixture.
Washing removes unbound components after target capture, reducing material that would otherwise remain mixed with the enriched fraction. The effectiveness of this stage follows from the difference between the target’s specific interaction with the immobilized ligand and the lack of binding by other constituents. A cleaner fraction supports subsequent analysis.
Release occurs when buffer conditions are changed or a competing molecule is added, because either intervention can weaken or displace the target’s interaction with the immobilized ligand. This step separates the captured molecule from the matrix after unwanted components have been removed. The released fraction can then support downstream biochemical analysis.
A typical workflow has four functional stages: introduce the sample to the binding matrix, allow the target to attach, wash away unbound material, and alter the buffer or add a competitor to release the target. Keeping capture, washing, and release conceptually separate helps identify where selectivity is created and where the enriched fraction is recovered.
Affinity Enrichment can concentrate several classes of biochemical targets, including proteins, antibodies, nucleic acids, and tagged biomolecules. The relevant requirement is the presence of a suitable specific binding interaction with the immobilized ligand. This flexibility makes the technique useful across different purification and characterization workflows.
In biochemistry, the enriched material can support more than purification. The technique is used for interaction studies, biomarker detection, and characterization of molecular complexes, as well as isolating targets for downstream analysis. These applications take advantage of selective recovery, making molecules or molecular complexes more accessible for focused biochemical examination.