Dithiothreitol
Dithiothreitol - Related Videos
03:39
Preparation of Formalin-Fixed Paraffin-Embedded Tumor Samples: A Protocol to Preserve Tissue and Cytology Samples from Non-small Cell Lung Cancer (NSCLC)
0 Views •
2023
This protocol describes the preparation of formalin-fixed paraffin-embedded tumor samples which are used to preserve tissue and cytology samples from non-small cell lung cancer...
08:01
LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics
0 Views •
Cited by 6 •
2017
Here we present a step-by-step protocol of the long-length electrostatic repulsion-hydrophilic interaction chromatography-tandem mass spectrometry (LERLIC-MS/MS) method. This is a novel methodology that enables for the first time quantification and characterization of the glutamine and asparagine deamidation isoforms by shotgun proteomics.
03:22
SDS-PAGE Based Extraction of Extracellular Vesicles Associated Proteins: A Procedure to Extract Proteins from EVs and Prepare Them for In-Gel Digestion
0 Views •
2023
In this video, we demonstrate the SDS-PAGE in-gel digestion method for protein extraction from extracellular vesicles or EVs. Once isolated, the obtained protein fragments can be used for proteomic analysis.
03:35
Assessing Viral Replication Capacity Using a Radiolabeled Reverse Transcriptase Assay
0 Views •
2026
Source: Claiborne, D. T. et al. A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses. J. Vis. Exp. (2014)This video demonstrates the assessment of viral replication capacity using a radiolabeled reverse transcriptase assay. Culture supernatants containing progeny virions are added to a reverse transcription mixture containing a radiolabeled nucleotide. The incorporated signal reflects viral output and replication...
09:39
Mapping Dysfunctional Protein-Protein Interactions in Disease
0 Views •
2025
Here, we present a protocol to enable the capture and identification of disease-specific protein-protein interactions from native cells and tissues using chemical probes and mass spectrometry. The resulting interaction datasets are analyzed through a dedicated web-based platform to reveal dynamic network dysfunctions and pathway alterations linked to disease.
FAQs
DTT transfers electrons through its two thiol groups, breaking the disulfide linkage and generating free sulfhydryl groups on the protein. During this reaction, DTT itself becomes oxidized into a disulfide form. This conversion explains why the reagent can maintain chemically reduced conditions while undergoing a defined change in its own molecular state.
Disulfide bonds can help maintain a protein's folded or linked structure. Reducing those bonds with DTT helps unfold disulfide-linked proteins and separates structural constraints that could influence analysis. As a result, researchers can examine proteins under standardized reduced conditions during electrophoresis and other assays, making the observed molecular behavior easier to interpret.
When disulfide bonds are converted to free sulfhydryl groups, researchers can examine proteins in a reduced chemical state rather than one maintained by those linkages. This is useful for investigating protein structure and enzyme activity, because the preparation preserves a specific redox condition that may influence how the target protein behaves during an assay.
DTT can be included during sample preparation, electrophoresis, and laboratory assays when researchers need proteins to remain reduced. Its role differs across these stages: preparation helps establish the desired chemical state, electrophoresis analyzes the treated protein, and assays evaluate molecular or enzymatic properties under conditions shaped by reduction.
By maintaining molecules in a chemically reduced state, DTT helps prevent disulfide-linked changes from altering the target protein during experimental handling and analysis. This supports more consistent examination of protein structure and activity. Its value is therefore procedural as well as chemical, particularly when reduction is important to the assay's interpretation.
DTT is primarily an experimental reagent that supports molecular investigations rather than a routine therapeutic treatment. In medical research, it can help examine redox regulation, enzyme activity, protein structure, and disease-related molecular changes. These applications use its reducing behavior to clarify biochemical mechanisms without implying that the reagent itself is a standard clinical intervention.