Digitonin Solubilization
Digitonin Solubilization - Related Videos
02:23
Visualizing the Effect of pH on Solubilization of the Influenza A Viral Core
0 Views •
2026
Source: Stauffer, S. et al., In Vitro Disassembly of Influenza A Virus Capsids by Gradient Centrifugation. J. Vis. Exp. (2016)This video demonstrates the effect of pH on the solubilization of influenza A viral cores. Gel electrophoresis and protein staining reveal the progressive disassembly of the core structure under acidic conditions.
10:43
Expression, Detergent Solubilization, and Purification of a Membrane Transporter, the MexB Multidrug Resistance Protein
0 Views •
Cited by 5 •
2010
In this protocol we demonstrate the expression, solubilization, and purification of a recombinantly expressed membrane protein, MexB, as a soluble protein detergent complex. MexB is a multidrug resistance membrane transporter from the opportunistic bacterial pathogen Pseudomonas aeruginosa.
03:05
Fluorescence-Detection Size-Exclusion Chromatography: A Technique to Identify the Integrity of Fluorescent Membrane Proteins upon Detergent Solubilization
0 Views •
2025
In this video, we demonstrate a fluorescence-detection size-exclusion chromatography technique to screen for green fluorescent protein (GFP)-fused membrane protein stability in Escherichia coli following detergent solubilization. Proteins exhibiting a symmetrical peak in the size-exclusion profile with little or no free GFP indicate purified proteins with minimum degradation and aggregation, which can subsequently be selected for structure-function analysis.
08:55
Expression, Solubilization, and Purification of Eukaryotic Borate Transporters
0 Views •
Cited by 5 •
2019
Here we present a protocol to express, solubilize, and purify several eukaryotic borate transporters with homology to the SLC4 transporter family using yeast. We also describe a chemical cross-linking assay to assess the purified homomeric proteins for multimeric assembly. These protocols can be adapted for other challenging membrane proteins.
13:06
Solubilization and Bio-conjugation of Quantum Dots and Bacterial Toxicity Assays by Growth Curve and Plate Count
0 Views •
Cited by 7 •
2012
Nanoparticles such as semiconductor quantum dots (QDs) can be used to create photoactivatable agents for anti-microbial or anti-cancer applications. This technique shows how to water-solubilize cadmium telluride (CdTe) QDs, conjugate them to an antibiotic, and perform a bacterial inhibition assay based upon growth curves and plate count.
FAQs
Digitonin binds cholesterol within lipid bilayers and forms digitonin-cholesterol complexes that disturb membrane organization. This interaction preferentially affects membranes containing sufficient cholesterol, allowing researchers to alter membrane permeability without necessarily disrupting every cellular membrane to the same extent. The resulting selectivity supports controlled access to cytosolic contents and membrane-associated components while potentially preserving intracellular structures under suitable conditions.
The main controlling variables are digitonin concentration, exposure time, temperature, and buffer conditions. Increasing or changing these parameters alters how extensively membranes are disrupted and how much cellular material becomes accessible. Researchers adjust the conditions according to whether they seek limited plasma-membrane permeabilization, cytosolic extraction, or more extensive membrane disruption for biochemical analysis.
Controlled disruption can provide access to selected cellular contents while helping preserve intracellular structures. This balance matters when researchers need to examine membrane-associated components, protein localization, or organelle function without treating the entire cell as a completely disorganized mixture. The quality of the resulting preparation therefore depends on matching the extent of permeabilization to the biological question.
The method can be adjusted to produce selective plasma-membrane permeabilization rather than uniformly disrupting all cellular membranes. Because digitonin acts through cholesterol-rich membrane regions, the concentration, exposure duration, temperature, and buffer collectively influence which cellular compartments become accessible. This distinction helps investigators study cytosolic material while retaining intracellular structures sufficiently for membrane organization or organelle-function studies.
A typical workflow begins by selecting digitonin conditions appropriate to the intended level of membrane disruption, including concentration, exposure time, temperature, and buffer. The treated cells can then provide access to cytosolic contents or membrane-associated components for downstream biochemical analysis. Researchers interpret the preparation according to how effectively the selected conditions balanced accessibility with structural preservation.
Researchers may select this approach for cell fractionation, cytosolic extraction, membrane-protein isolation, biochemical assays, or investigations of organelle function. It is especially useful when the experiment requires controlled access to cellular material rather than complete membrane destruction. The method can also support studies of protein localization and membrane organization by maintaining suitable intracellular structure during preparation.
Digitonin-treated cells can help reveal whether proteins or other components remain associated with membranes or become accessible after controlled permeabilization. By adjusting disruption conditions and examining the resulting cellular fractions or biochemical assay material, researchers can investigate membrane organization, protein localization, and compartment-specific accessibility. These outcomes connect membrane behavior with the distribution of intracellular components.