Np-40 Solubilization
Np-40 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
NP-40 interacts with membrane lipids and inserts into the lipid bilayer, weakening its organization. As the bilayer breaks apart, membrane proteins become associated with detergent molecules and can remain dispersed in the aqueous fraction. This mechanism allows researchers to recover membrane-associated proteins without relying on the more disruptive effects of harsher detergents.
Because NP-40 is nonionic, it generally disrupts lipid membranes while exerting less severe effects on protein structure than harsher detergents. Some biologically relevant protein-protein associations may therefore remain detectable after extraction. This property is useful when the goal is not only to measure a protein, but also to examine complexes or interactions involving membrane-associated components.
These variables determine the balance between membrane disruption and preservation of native protein relationships. Increasing detergent exposure can improve extraction, whereas overly strong or prolonged treatment may reduce the retention of biologically relevant complexes. Temperature also influences the extraction conditions. Researchers control all three factors to obtain sufficient solubilization without unnecessarily compromising protein structure or interactions.
A typical workflow applies NP-40 to cells or a cellular sample under controlled conditions, allowing the detergent to disrupt membranes and release membrane-associated components into an aqueous extract. The resulting preparation can then serve as material for antigen preparation, immunoprecipitation, or protein analysis. Exact detergent conditions are adjusted according to the desired balance between extraction efficiency and complex preservation.
The method is useful when investigators need access to host or pathogen membrane proteins for downstream study. Extracted material can support antigen preparation and immunoprecipitation, helping researchers examine target proteins or associated components. Its relatively gentle extraction behavior is especially relevant when membrane-associated interactions may contribute to immune recognition or infection-related protein organization.
NP-40 extracts can provide samples for techniques such as Western blotting, which supports detection and analysis of recovered proteins. The quality of the result depends on how well the extraction conditions disrupted membranes while retaining the target protein and, when relevant, its associated interactions. This makes controlled solubilization important for interpreting host or pathogen protein measurements.