Fplc System
Fplc System - Related Videos
10:39
Isolation of High-density Lipoproteins for Non-coding Small RNA Quantification
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Cited by 40 •
2016
This protocol describes the isolation and quantification of high-density lipoprotein small RNAs.
02:59
Anion-Exchange Chromatography-Based Protein Purification: A Separation Technique to Isolate a Protein of Interest From Dialyzed Bacterial Lysate Based on Net Charge
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2025
In this video, we demonstrate an anion-exchange chromatography technique to purify human calcium-binding protein S100A12 based on the net surface charge of the protein from an Escherichia coli culture.
03:45
Purification of Viral Integrase Using Affinity Chromatography
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2026
Source: Lopez Jr., M. A., et al. Detection and Removal of Nuclease Contamination During Purification of Recombinant Prototype Foamy Virus Integrase. J. Vis. Exp. (2017)This video demonstrates the purification of polyhistidine-tagged viral integrase using nickel affinity chromatography. The protocol uses imidazole gradient elution to selectively isolate the integrase based on its affinity for nickel-charged resin.
12:03
Expression, Isolation, and Purification of Soluble and Insoluble Biotinylated Proteins for Nerve Tissue Regeneration
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Cited by 12 •
2014
Developing biotinylatable fusion proteins has many potential applications in various fields of research. Recombinant protein engineering is a straight forward procedure that is cost-effective, providing high yields of custom-designed proteins.
07:50
Detection and Removal of Nuclease Contamination During Purification of Recombinant Prototype Foamy Virus Integrase
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Cited by 10 •
2017
Recombinant prototype foamy virus integrase protein is often contaminated with a bacterial nuclease during purification. This method identifies nuclease contamination and removes it from the final preparation of the enzyme.
FAQs
Separation occurs because biomolecules interact with the chromatography column differently according to size, charge, hydrophobicity, or affinity. Those differences alter how quickly each molecule moves with the buffer, producing distinct elution times. The resulting separation allows a mixed sample to be resolved into fractions that can be collected for later protein purification or analysis.
Buffer conditions can be changed as a gradient, while pumps maintain a controlled liquid-flow rate through the column. Together, these features help expose differences in molecular behavior and produce a more controlled elution pattern. Automated gradient control also supports reproducible separations, which is important when researchers compare protein fractions or repeat purification workflows.
The detector tracks signals such as ultraviolet absorbance as material leaves the column. Changes in that signal indicate when biomolecules elute and help identify portions of the run suitable for fraction collection. This monitoring connects the instrument’s separation behavior with an observable output, supporting both purification and analysis of the resulting biomolecular fractions.
Automated pumping, gradient control, detection, and fraction collection reduce variation between runs. This consistency improves reproducibility while controlled liquid handling helps preserve sensitive biological samples. In practice, the combination is useful when a study requires comparable protein preparations, repeated fractionation, or dependable examination of biomolecular interactions rather than a single manually managed separation.
A typical workflow places the sample onto a chromatography column, drives buffer through it with regulated pumps, monitors the elution signal, and collects selected fractions. The column separates components according to their molecular properties, while the detector provides a record of when material emerges. Collected fractions can then support protein purification or downstream biological analysis.
The system is useful when researchers need to separate, purify, or fractionate proteins and other biomolecules before examining their properties. It can support studies of protein structure, enzyme activity, biomolecular interactions, and recombinant products. Fraction collection is especially valuable because it preserves separated portions for subsequent biological characterization instead of treating the mixture as one sample.