Pbmc Separation
Pbmc Separation - Related Videos
02:26
A Technique to Generate a Human PBMC-Engrafted Humanized Xenograft Mouse Model
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2025
This video demonstrates an assay to generate a humanized xenograft mouse model. An immunocompromised mouse is injected with cyclophosphamide to cause immune cell depletion. Patient-derived tumor cells mixed with human peripheral blood mononuclear cells (PBMCs) in a hydrogel are subcutaneously injected into the mouse, forming a tumor with infiltrating T cells.
08:02
Separation of Mixtures via Precipitation
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2023
Source: Laboratory of Dr. Ana J. García-Sáez — University of Tübingen Most samples of interest are mixtures of many different components. Sample preparation, a key step in the analytical process, removes interferences that may affect the analysis. As such, developing separation techniques is an important endeavor not just in academia, but also in industry. One way to separate mixtures is to use their solubility properties. In this short paper, we will deal with aqueous solutions. The...
07:29
Separating Protein with SDS-PAGE
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2023
Sodium Dodecyl Sulfate Poly-Acrylamide Gel Electrophoresis, or SDS-PAGE, is a widely-used technique for separating mixtures of proteins based on their size and nothing else. SDS, an anionic detergent, is used to produce an even charge across the length of proteins that have been linearized. By first loading them into a gel made of polyacrylamide and then applying an electric field to the gel, SDS-coated proteins are then separated. The electric field acts as the driving force, drawing the SDS...
08:17
Electrophoretic Separation of Proteins
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Cited by 29 •
2008
In this video, we demonstrate a method for electrophoretic separation of proteins using poly-acrylimide gel electrophoresis (PAGE).
11:40
A Co-culture Method to Investigate the Crosstalk Between X-ray Irradiated Caco-2 Cells and PBMC
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Cited by 7 •
2018
We present a protocol to investigate the crosstalk between X-ray-irradiated Caco-2 and peripheral blood mononuclear cells (PBMC). The protocol starts with Caco-2 irradiation and set-up of the co-culture with PBMC; subsequently, trans-epithelial electrical resistance is measured regularly over 48 h and western blot performed in both Caco-2 and PBMC.
FAQs
The separation relies on differences in cell density rather than immune-cell identity. After blood is diluted and layered over a density-gradient medium, centrifugation produces distinct regions: PBMCs remain at the plasma–gradient interface, whereas denser erythrocytes and granulocytes move below the gradient. This physical separation concentrates lymphocytes and monocytes for downstream investigation.
Ficoll provides the density-gradient medium that enables blood components to distribute into separate layers during centrifugation. Its position beneath the diluted blood creates an interface where the less dense mononuclear cells collect, while erythrocytes and granulocytes sediment farther downward. The resulting band can be recovered for washing and subsequent cellular or molecular analysis.
The recovered fraction is enriched mainly for lymphocytes and monocytes, the mononuclear cells most relevant to many immune studies. Erythrocytes and granulocytes are separated below the gradient rather than collected with this fraction. Consequently, the preparation provides a focused starting population for examining immune-cell behavior, infection-related responses, or inflammatory processes.
Once the PBMC layer is recovered from the plasma–gradient interface, the cells can be washed to remove residual gradient medium and other separated material. Researchers may then count the cells, culture them, or prepare them for flow cytometry and molecular assays. These steps convert the isolated fraction into a defined input for experiments and measurements.
This preparation is useful when investigators need access to circulating mononuclear immune cells for studying host–pathogen interactions, inflammation, vaccine responses, or infectious-disease biomarkers. Because lymphocytes and monocytes are recovered as a workable cell fraction, the same starting material can support functional studies, cellular profiling, or molecular analysis across diverse immune and infection experiments.
Separated PBMCs can provide material for several complementary outcomes. Cell counting measures the recovered population, culture enables investigation of cellular function, flow cytometry supports analysis of immune-cell characteristics, and molecular assays examine biological signals. Together, these approaches can help evaluate immune responses, infection-associated changes, vaccine effects, and candidate biomarkers in the studied samples.