Dsp Cross-linker
Dsp Cross-linker - Related Videos
12:30
Vascular Gene Transfer from Metallic Stent Surfaces Using Adenoviral Vectors Tethered through Hydrolysable Cross-linkers
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Cited by 12 •
2014
These studies report on reversible attachment of adenoviral gene vectors to coatless metal surfaces of stents and model mesh disks. Sustained release of transduction-competent viral particles contingent upon hydrolysis of cross-linkers used for vector immobilization results in a durable site-specific transgene expression in vascular cells and in stented arteries.
10:34
Identification of Sleeping Beauty Transposon Insertions in Solid Tumors using Linker-mediated PCR
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Cited by 3 •
2013
A method of identifying unknown drivers of carcinogenesis using an unbiased approach is described. The method uses the Sleeping Beauty transposon as a random mutagen directed to specific tissues. Genomic mapping of transposon insertions that drive tumor formation identifies novel oncogenes and tumor suppressor...
01:09
Cytoskeletal Linker Proteins - Plakins
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2023
Plakins are large proteins with binding domains for microtubules, microfilaments, intermediate filaments, and membrane-associated protein complexes at cell junctions. Plakin functions are evolutionarily conserved and are primarily involved in organizing the different components of the cytoskeleton by crosslinking them to each other and connecting them to the cell-matrix and cell adhesion complexes. They are also known to interact with signal transducers, serve as scaffolds for signaling...
14:40
Expression Analysis of Mammalian Linker-histone Subtypes
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Cited by 22 •
2012
We describe a set of assays to analyze expression levels of H1 linker histones. mRNA of individual H1 genes are quantitatively measured by random primer based reverse transcription followed by real-time PCR, whereas protein quantification of H1 histones is achieved by HPLC analysis.
10:50
Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography
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Cited by 37 •
2010
The cell permeable crosslinker DSP [dithiobis-(succinimidyl propionate)] stabilizes transient and labile interactions in vivo, which allows their isolation using stringent protein complex purification techniques. Here we present a technique for crosslinking cells grown in culture followed by isolation of protein complexes by immunoprecipitation.
FAQs
The two N-hydroxysuccinimide ester groups react with primary amines, including lysine side chains, and form covalent amide bonds. Because the reactive groups are connected by a spacer, the reagent can stabilize biomolecules that are close enough for both ends to react. This converts a potentially transient association into a chemically preserved connection for later analysis.
The disulfide bridge makes the resulting cross-links reversible under reducing conditions. Researchers can first stabilize a molecular complex, then cleave the linkage during analysis when reduction is applied. This combination preserves interaction information while allowing the experimenter to distinguish or release components that had been covalently connected.
Membrane permeability allows DSP cross-linker to access protein interactions within cells rather than limiting stabilization to material exposed after lysis. That property is useful when molecular organization may change during sample disruption. By acting before or during lysis-related handling, the reagent can help retain interaction patterns from complex biological samples.
Covalent stabilization reduces the chance that weak or short-lived associations will dissociate during cell lysis and subsequent sample handling. The reagent therefore preserves proximity relationships that might otherwise be lost before detection. This is especially relevant when researchers want to examine protein complexes or molecular organization rather than only the most stable interactions.
A supported workflow uses DSP cross-linker to stabilize cellular or biochemical associations, followed by cell lysis or other sample handling that would otherwise disrupt them. The preserved material can then be examined through immunoprecipitation and electrophoresis. These steps connect chemical stabilization with separation and detection of the resulting protein complexes.
After interactions have been chemically preserved, immunoprecipitation can be used to isolate complexes associated with a selected protein or molecular target. The cross-linking step helps retain associated partners during lysis and purification. Analysis of the recovered material can therefore reveal interaction partners or complex composition that might be underestimated without stabilization.
Electrophoresis provides a way to examine the molecular species produced after cross-linking and sample preparation. Researchers can compare material containing preserved complexes with material treated under reducing conditions, where the disulfide-containing linkage can be cleaved. These patterns help assess complex organization and the presence of cross-linked protein assemblies.
DSP cross-linker supports biochemical studies that require both preservation and later interrogation of protein assemblies. Its membrane permeability, amine-reactive ends, and reducible disulfide bridge allow researchers to capture interactions in complex samples, analyze them by immunoprecipitation or electrophoresis, and examine how proteins are arranged within larger molecular structures.