Nitrocellulose Membrane Overlay

Nitrocellulose Membrane Overlay is a laboratory technique that transfers and immobilizes proteins or other biological components from a sample onto a porous nitrocellulose membrane for detection. When the membrane contacts a cell culture, microbial colony, or protein-containing surface, biomolecules bind to the membrane through hydrophobic and electrostatic interactions; researchers can then probe the captured material with specific antibodies or other detection reagents. In immunology and infection research, this approach supports antigen screening, identification of pathogen-associated proteins, and analysis of host-pathogen interactions. Its simple workflow helps link molecular composition to immune recognition and can guide diagnostic or mechanistic studies.

Nitrocellulose Membrane Overlay - Related Videos

Research

JoVE Journal - Biology

Protein Membrane Overlay Assay: A Protocol to Test Interaction Between Soluble and Insoluble Proteins in vitro

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Cited by 6 •

2011

Testing protein-protein interaction is indispensable for dissection of protein functionality. Here, we introduce an in vitro protein-protein binding assay to probe a membrane-immobilized protein with a soluble protein. This assay provides a reliable method to test interaction between an insoluble protein and a protein in solution.

Quantification of Bacterial Histidine Kinase Autophosphorylation Using a Nitrocellulose Binding Assay

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Cited by 3 •

2017

We report and demonstrate an optimized nitrocellulose binding assay that can be used to quantify autophosphorylation of purified bacterial histidine kinases. Our method has several advantages over traditional SDS-PAGE based techniques, providing a valuable alternative for characterizing these important proteins.

Evaluation of a Liquid Overlay System for Viral Plaque Assays

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2026

Source: Baer, A. and Kehn-Hall, K. Viral Concentration Determination Through Plaque Assays: Using Traditional and Novel Overlay Systems. J. Vis. Exp. (2014).This video demonstrates the use of a liquid microcrystalline cellulose-based overlay in a plaque-based assay by comparing its effectiveness in viral plaque formation with that of a traditional semi-solid agarose overlay.

Research

JoVE Journal - Immunology and Infection
Free Sample

Viral Concentration Determination Through Plaque Assays: Using Traditional and Novel Overlay Systems

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Cited by 466 •

2014

Plaque assays are the gold standard for viral quantification, utilizing entrapping overlays on host cellular monolayers to determine viral titers. While various semisolid overlays have traditionally been used, here we demonstrate plaque techniques comparing semisolid overlays to a novel liquid microcrystalline cellulose among several families of viruses.

Use of the Soft-agar Overlay Technique to Screen for Bacterially Produced Inhibitory Compounds

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Cited by 141 •

2017

We describe a simple method for screening bacterial cultures for the production of compounds inhibitory towards other bacteria.

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