Cell Purification
Cell Purification - Related Videos
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.
03:22
Production and Purification of Recombinant Antibodies from Mammalian Cells
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2025
The video explains the process of producing antibodies in mammalian host cells, from gene expression to secretion. It involves transcription, translation, signal recognition, correct folding, and eventual release into the extracellular medium via secretory vesicles.
11:30
Tandem Affinity Purification of Protein Complexes from Eukaryotic Cells
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Cited by 7 •
2017
We describe here a novel, robust, and efficient tandem affinity purification (TAP) method for the expression, isolation, and characterization of protein complexes from eukaryotic cells. This protocol could be utilized for the biochemical characterization of discrete complexes as well as the identification of novel interactors and post-translational modifications that regulate their function.
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.
FAQs
It separates cells according to buoyancy, so populations with different densities form distinct layers after centrifugation. Researchers can collect the layer containing the desired cells while excluding material that settles or remains elsewhere. This approach is useful when cell density provides a practical distinction between populations before immunological or infection-related assays.
Both approaches use defined surface markers to identify cells of interest. Antibody-based magnetic sorting enriches cells according to marker recognition, whereas flow cytometry provides marker-based selection during analysis and sorting. Their specificity allows researchers to focus on populations such as T cells, B cells, or monocytes rather than analyzing all cells together.
A more uniform cell population makes measured responses easier to attribute to the cells being studied. This improves the accuracy and reproducibility of molecular and functional assays, including experiments examining immune activation, pathogen susceptibility, host-pathogen interactions, or therapeutic responses. Without adequate enrichment, signals from unwanted populations can complicate interpretation.
The choice depends on which differences distinguish the target population from surrounding cells. Density and size support physical separation approaches, while surface markers support antibody-based magnetic sorting or flow cytometry. Biological properties may also provide a useful basis for selection. Matching the strategy to the available distinguishing feature helps enrich the intended cells.
Researchers can purify a selected population before testing its structure, function, or response to an experimental condition. Peripheral blood mononuclear cells, T cells, B cells, monocytes, and infected cell populations may each serve as starting or enriched material. The resulting preparation supports more focused molecular and functional measurements than a heterogeneous sample.
Purified populations enable focused studies of immune activation, pathogen susceptibility, host-pathogen interactions, and therapeutic responses. For example, separating T cells, B cells, or monocytes allows investigators to examine responses associated with a defined cellular population, while enriching infected cells supports analysis of infection-related behavior without relying solely on measurements from mixed samples.