Fluorescent Probe Conjugation
Fluorescent Probe Conjugation - Related Videos
04:02
An Assay to Measure the Accumulation of a Fluorescent Antibiotic Probe in Bacterial Cells
0 Views •
2025
This video demonstrates an assay to measure the accumulation of a fluorescent antibiotic probe in bacterial cells. Exposure to an efflux pump inhibitor disrupts the transmembrane proton gradient in bacterial cells, upon which a fluorophore-conjugated antibiotic is added. Inhibition of efflux pumps due to the disrupted proton gradient causes accumulation of the antibiotic inside the bacteria. Bacterial cells are lysed, and the fluorescence intensity is measured to confirm antibiotic accumulation.
10:18
Making Conjugation-induced Fluorescent PEGylated Virus-like Particles by Dibromomaleimide-disulfide Chemistry
0 Views •
Cited by 12 •
2018
Here, we present a procedure to fluorescently functionalize the disulfides on Qβ VLP with dibromomaleimide. We describe Qβ expression and purification, the synthesis of dibromomaleimide-functionalized molecules, and the conjugation reaction between dibromomaleimide and Qβ. The resulting yellow fluorescent conjugated particle can be used as a fluorescence probe inside cells.
07:14
Assessing Phagocytic Clearance of Cell Death in Experimental Stroke by Ligatable Fluorescent Probes
0 Views •
Cited by 4 •
2014
We present a new fluorescence technique for selective in situ labeling of active phagocytic cells, which clear off cell corpses in stroke. The approach is important for assessing brain reaction to ischemia because only a small proportion of phagocytes present in ischemic brain participate in clearance of cell death.
02:50
Conjugated Proteins
0 Views •
2020
Simple proteins and protein complexes contain only amino acids. In contrast, many other proteins, called conjugated proteins, covalently bond with non-protein moieties. Nucleoproteins are protein complexes that contain nucleic acids, categorized as deoxyribonucleoproteins (DNPs) or ribonucleoproteins (RNPs) respectively. The nucleosome is a typical example of a DNP where nuclear DNA is associated with histone proteins. The major antigen for the Covid-19 virus SARS-CoV is an RNP that is critical...
10:59
Determination of Lipid Raft Partitioning of Fluorescently-tagged Probes in Living Cells by Fluorescence Correlation Spectroscopy (FCS)
0 Views •
Cited by 9 •
2012
A technique to probe the lipid raft partitioning of fluorescent proteins at the plasma membrane of living cells is described. It takes advantage of the disparity in diffusion times of proteins located inside or outside of lipid rafts. Acquisition can be performed dynamically in control conditions or after drug addition.
FAQs
The reaction depends on matching chemical groups: an activated fluorophore must encounter a compatible functional group on the selected biomolecule. This pairing creates a covalent attachment rather than a temporary association, allowing the fluorescent signal to remain linked to the protein, peptide, nucleic acid, or other target during subsequent observation and analysis.
pH, reactant concentration, and reaction time are central control variables. They determine how effectively the activated fluorophore encounters and reacts with functional groups on the target biomolecule. Maintaining controlled conditions helps produce a usable labeled product, while poor control can reduce the effectiveness or consistency of the conjugation process.
Unreacted fluorescent probe must be separated from the conjugated biomolecule before analysis. Otherwise, free probe can contribute fluorescence that does not represent the target, complicating interpretation of microscopy, spectroscopy, or flow-based measurements. Removing it makes the observed signal more closely associated with the labeled biomolecule and its behavior.
Mixing places the components together, whereas conjugation creates a covalent link between the fluorophore and a compatible group on the biomolecule. That chemical attachment allows the label to follow the target during measurements of location, movement, or interactions. The distinction is important when researchers need the fluorescence to report on the biomolecule rather than free probe.
A typical workflow selects a biomolecule and a compatible activated fluorophore, combines them under controlled pH, concentration, and reaction-time conditions, and then removes probe that did not react. The resulting labeled biomolecule can be examined using fluorescence microscopy, spectroscopy, or flow-based analysis, depending on the information required.
Fluorescent conjugates can make a biomolecule’s location, movement, or interactions observable. Microscopy is suited to examining where labeled material appears, while spectroscopy or flow-based analysis can support measurements across samples or populations. The chosen readout therefore connects the same labeling strategy to different questions about biomolecular behavior.
In bioengineering, the technique supports biosensor development, assay design, and molecular tracking. It also helps researchers evaluate engineered cells, materials, and therapeutic delivery systems by providing a fluorescent handle for observation or flow-based analysis. These uses connect molecular labeling with the design and assessment of engineered biological systems.