Proteinase K
Proteinase K - Related Videos
09:45
Detection of Copy Number Alterations Using Single Cell Sequencing
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Cited by 9 •
2017
Single cell sequencing is an increasingly popular and accessible tool for addressing genomic changes at high resolution. We provide a protocol that uses single cell sequencing to identify copy number alterations in single cells.
08:38
Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery
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Cited by 29 •
2018
Here, a protocol for medium- to high-throughput analysis of protein phosphorylation events at the cellular level is presented. Phospho flow cytometry is a powerful approach to characterize signaling aberrations, identify and validate biomarkers, and assess pharmacodynamics.
14:44
Harmonic Radar Tags for Insect Tracking: Lightweight, Low-cost, and Accessible
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Cited by 2 •
2025
This protocol presents the methods for creating, attaching, and tracking harmonic radar tags for insects.
11:41
Assessing Transmissible Spongiform Encephalopathy Species Barriers with an In Vitro Prion Protein Conversion Assay
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2015
Measuring the barrier to the interspecies transmission of prion diseases is challenging and typically involves animal challenges or biochemical assays. Here, we present an in vitro prion protein conversion assay with the ability to predict species barriers.
02:23
Obtaining Viral RNA for Real-Time Quantitative Polymerase Chain Reaction (qPCR) Analysis
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2026
Source: Suzuki, Y., et al. Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction. J. Vis. Exp. (2018).This video demonstrates the procedure for preparing viral samples for quantitative PCR (Polymerase chain reaction) analysis. Viral samples and RNA standards are serially diluted, treated with a proteinase K buffer, and incubated under controlled temperatures to release RNA. The processed RNA is then stabilized and stored for...
FAQs
Its broad activity comes from its ability to cleave peptide bonds in both native proteins, which retain their folded structures, and denatured proteins, whose structures have been disrupted. This substrate range allows the enzyme to attack diverse cellular proteins rather than targeting only one protein type, making it useful for comprehensive protein removal during molecular biology workflows.
Proteinase K retains activity under conditions that disrupt or modify protein structure, including the presence of detergents and denaturants. That stability is important because these reagents can help expose cellular proteins while the enzyme continues digesting them. The combination supports more thorough breakdown of protein contaminants during nucleic acid purification.
Digesting both forms broadens the range of contaminants that can be removed from a biological sample. Folded cellular proteins may be accessible to the enzyme in their native state, while denaturation can expose additional peptide bonds for cleavage. This flexibility improves the removal of protein material before DNA or RNA is used in downstream analyses.
The enzyme removes proteins that can remain associated with nucleic acids, including histones, which bind DNA, and nucleases, which can degrade DNA or RNA. By digesting these components, treatment helps release nucleic acids from cellular material and reduces protein-related interference or degradation, improving their suitability for subsequent molecular analyses.
Within a purification workflow, Proteinase K is used to digest cellular proteins and other protein contaminants while nucleic acids are being recovered. Its activity helps break down structural proteins, histones, and nucleases, supporting nucleic acid release and protection. The resulting preparation can then be directed toward analyses that require usable DNA or RNA.
Proteinase K treatment supports downstream applications identified in the source material, including PCR, sequencing, and genotyping. These analyses depend on recovered nucleic acids that are sufficiently free of protein contaminants and nucleases. By improving nucleic acid release and protection during purification, the treatment helps prepare samples for these molecular biology procedures.
Its broad-spectrum digestion allows Proteinase K to address many different proteins in a cellular sample rather than requiring a separate enzyme for each contaminant. It can also function in the presence of detergents and denaturants, giving the purification process flexibility when cellular material must be disrupted and protein components removed before nucleic acid analysis.