Reverse Crosslinking
Reverse Crosslinking - 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.
10:10
Reverse Total Shoulder Arthroplasty
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Cited by 5 •
2011
Reverse total shoulder arthroplasty is indicated for the treatment of conditions that cannot be treated with conventional arthroplasty or other procedures. These primarily include degenerative and incapacitating conditions with irreparable rotator cuff and loss of the normal biomechanical coupling of the shoulder.
09:39
Creation of Reversible Cholestatic Rat Model
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2011
Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model.
FAQs
Temperature and incubation time determine how effectively crosslinker-derived chemical bonds are cleaved while preserving the material needed for analysis. Insufficient treatment may leave biomolecules attached to proteins, lowering recovery and accessibility. Excessive or poorly controlled treatment can damage samples. Researchers therefore adjust these variables to balance bond reversal with reliable recovery for sequencing, amplification, or electrophoresis.
These treatments promote different aspects of bond reversal and sample release. Heat can support cleavage of chemical links, detergents can disrupt protein-associated complexes, and selected chemical conditions can help break crosslinker-derived bonds. Their use makes DNA, RNA, or proteins more accessible during purification and analysis, although the appropriate combination depends on the sample and downstream assay.
Residual crosslinks can keep nucleic acids associated with proteins or other biomolecules, limiting their recovery during purification and reducing access for later analysis. In chromatin or RNA-protein studies, this may weaken detection of binding sites or molecular associations. Completing the reversal under controlled conditions helps produce material suitable for amplification, sequencing, or electrophoretic analysis.
A typical workflow applies heat, detergent, or compatible chemical treatment to the crosslinked sample, followed by conditions that allow the bonds to reverse during incubation. The released DNA, RNA, or proteins are then recovered through purification and prepared for the selected assay. Temperature, reaction conditions, and incubation duration should be controlled and optimized for the downstream measurement.
In chromatin immunoprecipitation, reversal helps release nucleic acids from protein-associated complexes so researchers can examine binding sites and gene regulation. RNA-protein interaction studies use the same general strategy to recover RNA or associated material after fixed molecular associations have been analyzed. It is also relevant when preparing formaldehyde-fixed samples for downstream molecular characterization.
Successful treatment can improve recovery of DNA, RNA, or proteins and make them more suitable for downstream sequencing, amplification, or electrophoresis. In biology, these measurements can support analysis of molecular associations, protein binding sites, and gene-regulatory relationships. The quality of the result depends on achieving sufficient bond reversal without compromising the recovered sample.