Dsp Crosslinker
Dsp Crosslinker - Related Videos
03:47
Crosslinking Neuronal Surface Proteins in the Mouse Brain using a Chemical Crosslinking Assay
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2025
This video demonstrates the crosslinking of neuronal surface proteins using Bis-sulfosuccinimidyl suberate or BS3. Isolated and chilled mouse brain is sectioned coronally. The desired region is extracted from the sections, minced, and treated with BS3. BS3 crosslinks surface proteins like GABA receptors, leaving internal proteins unaltered. The reaction is quenched with glycine before processing the samples for further analysis.
11:15
Nanosponge Tunability in Size and Crosslinking Density
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Cited by 1 •
2017
This article describes a process for tuning the size and crosslinking density of covalently crosslinked nanoparticles from linear polyesters containing pendant functionality. By tailoring synthesis parameters (polymer molecular weight, pendant functionality incorporation, and crosslinker equivalents), a desired nanoparticle size and crosslinking density can be achieved for drug delivery applications.
09:06
Preparation of DNA-crosslinked Polyacrylamide Hydrogels
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Cited by 5 •
2014
Our laboratory has developed DNA-crosslinked polyacrylamide hydrogels, a dynamic hydrogel system, to better understand the effects of modulating tissue stiffness on cell function. Here, we provide schematics, descriptions, and protocols to prepare these hydrogels.
05:07
An Enhanced Crosslinking Immunoprecipitation Method for Efficient Identification of Protein-bound RNA in Mouse Testes
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2025
This video demonstrates the enhanced crosslinking immunoprecipitation (eCLIP) method for identifying RNA-binding proteins in mouse testis. eCLIP enables the precise identification of binding sites where RNA-binding proteins (RBPs) interact with their target RNAs.
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 N-hydroxysuccinimide ester groups react with accessible primary amines, including those on lysine residues, on molecules positioned near one another. This reaction converts a noncovalent association into a covalent connection through the spacer. Stabilizing these neighboring molecules helps preserve protein-protein interactions and other biomolecular assemblies for subsequent purification and analytical workflows.
The disulfide spacer creates an analytical release point. After crosslinked material has been isolated or examined, appropriate reducing conditions can reverse the linkage, allowing the associated components to be analyzed separately. This feature supports workflows that require both stabilization of an interaction in the original biological sample and later characterization of the individual proteins or other biomolecules.
Crosslink formation depends on whether reactive primary amines are accessible and positioned near a partnering molecule. Amines that are unavailable for reaction cannot efficiently participate, while nearby accessible groups can connect associated components. Consequently, the reagent reports molecular proximity within the sample rather than simply labeling every protein present, which helps preserve information about biological organization.
A cleavable crosslink can stabilize an assembly during biochemical processing and then be reversed under appropriate reducing conditions. That reversibility allows linked components to be separated for analysis instead of remaining permanently connected. The design therefore combines structural preservation with the option to examine the participating proteins or other biomolecules individually.
A biological sample can be treated to stabilize nearby interacting components before a downstream workflow such as biochemical purification, immunoprecipitation, or interaction-proteomics analysis. The crosslinked material preserves associations that might otherwise be disrupted during processing. If separate component analysis is needed, the disulfide-containing linkage can subsequently be reversed under appropriate reducing conditions.
The approach can support studies of protein-protein interactions, multiprotein complexes, membrane-associated assemblies, and cellular organization. By stabilizing associated molecules in biological samples, it helps researchers examine how components are connected within larger assemblies. Its compatibility with purification and immunoprecipitation workflows also makes it useful for isolating interaction-related material for further analysis.
In interaction-proteomics workflows, stabilization can preserve molecular associations long enough for linked material to be processed and examined. The resulting analysis can help identify components that were associated within a biological sample, while reduction of the disulfide linkage can permit separate examination of those components. This connects interaction mapping with follow-up characterization of individual molecules.