Stage Tip Purification
Stage Tip Purification - Related Videos
05:43
A STAGE Tip Procedure for Desalting and Purification of Peptides
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2026
Source:Tia Rizakos1, Jennifer Geddes-McAlister11Molecular and Cellular Biology Department, University of Guelph.This video demonstrates peptide purification through the STAGE Tip procedure. A miniature column containing resin beads bonded to long hydrocarbon chains facilitates hydrophobic interaction-based purification. Upon loading a pre-digested peptide sample into the column, the peptide fragments bind to the resin beads, allowing impurities to pass through. An elution buffer that disrupts...
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
06:30
Gel Purification
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2023
Gel purification is used to recover DNA fragments after electrophoretic separation. DNA recovery from an agarose gel includes three basic steps: binding, washing and eluting from a silica column. DNA is believed to bind to silica in the presence of high salt via a salt bridge. Following binding, DNA is washed of impurities and eluted under low salt conditions disrupting this interaction. This video goes through a step-by-step, generalized procedure for cutting out a band from the gel, gel...
11:22
GST-His purification: A Two-step Affinity Purification Protocol Yielding Full-length Purified Proteins
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Cited by 18 •
2013
In the present protocol, we demonstrate a highly efficient and cost-effective small-scale protein purification method, which allows purification of recombinant proteins by uniquely combining a cleavable GST-tag and a small His-tag.
01:22
Stages of General Anesthesia
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2024
Various sedation levels offer significant advantages in facilitating procedural interventions for patients undergoing medical or invasive surgical procedures. These levels span from anxiolysis to general anesthesia, providing a spectrum of sedative effects to cater to specific patient needs. Anxiolysis reduces anxiety and is achieved through minimal sedation, enabling patients to remain awake and responsive while feeling more at ease during the procedure. This level can benefit minor...
FAQs
Analyte retention is controlled by the defined solvent conditions applied when the sample contacts the chromatographic sorbent. These conditions determine whether target peptides or proteins bind strongly enough for contaminants to be removed later. Because binding depends on this chemical environment, consistent solvent conditions are important for efficient cleanup and reproducible recovery across biochemical samples.
Washing removes salts, detergents, and other contaminants that remain after analytes bind to the sorbent. This selective cleanup improves the chemical quality of the retained peptide or protein fraction without discarding the target molecules. Removing these substances is especially important before chromatography or mass spectrometry, where sample composition affects compatibility and detection.
Elution releases the bound target molecules from the chromatographic material after unwanted components have been washed away. The resulting fraction contains the cleaned analytes in a form suitable for downstream analysis, while the overall microscale format supports concentration as well as purification. Effective elution therefore determines whether the prepared sample can be recovered for subsequent biochemical measurements.
Packing chromatographic material into a pipette tip reduces the amount of sorbent, sample, and solvent needed for processing. This format suits limited biochemical samples and enables purification within small-scale workflows rather than requiring larger extraction setups. The reduced resource requirement is particularly useful in proteomics, where many samples may need consistent preparation before analytical measurements.
A typical workflow applies the biochemical sample to the packed chromatographic material under conditions that promote analyte binding. The sorbent is then washed to remove salts, detergents, and related contaminants, followed by elution of the retained target molecules. Keeping these stages distinct supports cleanup and concentration while preparing the sample for chromatography or mass spectrometry.
Researchers use this approach when peptide or protein samples require cleanup and concentration before chromatography or mass spectrometry. It is also valuable for microscale and proteomics workflows because it uses small sample and solvent volumes. By improving sample compatibility and supporting reproducible preparation, the method helps produce biochemical fractions that are more suitable for downstream detection and analysis.