Nitrocellulose Filter Binding
Nitrocellulose Filter Binding - Related Videos
09:17
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.
01:27
Passive Filters
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2024
Passive filters are utilized to shape the frequency spectrum of signals across a diverse array of applications. These filters, using only passive elements like resistors (R), inductors (L), and capacitors (C), are capable of selectively allowing or blocking certain frequency ranges without the need for external power sources. Low-Pass Filters Low-pass filters are designed to transmit signals with frequencies lower than the cutoff frequency, ωc, and attenuate those above it. The cutoff frequency...
09:32
Evaluating the Effect of Coated Filters and Pre-Filters on the Infectivity and Concentration of Aerosolized Phi6 Virus
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2026
Here, we present a study to evaluate the anti-viral efficacy of copper- and silver-based surface coatings on air filters. Using a bioaerosol chamber and Phi6 bacteriophage, this method quantifies viral inactivation and mechanical filtration efficiency for self-sanitizing indoor air technologies.
01:25
Active Filters
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2025
Active filters are electronic circuits that use operational amplifiers (op-amps), resistors, and capacitors to filter out unwanted frequency components from a signal. A first-order low-pass active filter is designed to pass signals with a frequency lower than a certain cutoff frequency and attenuate frequencies higher than that cutoff frequency. The transfer function for a first-order low-pass active filter is: Where Af is the gain at low frequencies also known as the dc gain, s is the...
06:30
SELEX-Based In Vitro Binding Assay to Identify RNA-Protein Interactions
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2025
In this video, we describe the systematic evolution of ligands by the exponential enrichment (SELEX) method to identify specific RNA-binding sequences for a target protein of interest. The protein is incubated with a large pool of randomized RNA sequences, and the protein-binding RNA sequences are isolated, PCR-amplified, and sequenced to identify their protein-binding sites.
FAQs
The membrane provides the assay’s physical discrimination: protein and protein-containing complexes remain associated with nitrocellulose, whereas nucleic acid that has not formed such an association passes through. Consequently, the retained fraction represents nucleic acid captured through protein binding rather than the total nucleic acid supplied, allowing binding to be quantified.
Binding affinity is inferred from how much nucleic acid remains associated under the chosen assay conditions, while specificity is assessed by comparing interactions involving different DNA or RNA sequences. Changing sequence or experimental conditions can therefore reveal whether a protein recognizes particular nucleic-acid features and how strongly that recognition is maintained.
Because the readout depends on whether nucleic acid is retained with a protein or protein complex, the approach can examine interactions involving either DNA or RNA. This flexibility supports comparisons between DNA-binding and RNA-binding proteins, while sequence comparisons help determine whether recognition differs among the nucleic-acid targets tested.
At a basic level, the workflow has three linked stages: incubate the protein or protein complex with DNA or RNA, pass the mixture through a nitrocellulose membrane, and measure the nucleic acid retained on the membrane. Keeping these stages conceptually separate helps connect the measured signal to the interaction under investigation.
A labeled probe converts retained nucleic acid into a measurable readout. Comparing that signal across different nucleic-acid sequences or experimental conditions can indicate changes in binding affinity or specificity. The result is therefore not merely evidence that material remained on the membrane; it supports quantitative or comparative analysis of the interaction.
In genetics research, the assay is particularly useful for examining transcription factors, DNA-binding proteins, RNA-binding proteins, and other regulatory interactions. These applications connect a biochemical binding measurement with questions about gene regulation and molecular recognition, including whether a protein interacts selectively with a DNA or RNA sequence involved in regulation.