Creative Proteomics uses a variety of primary cell separation methods including enzymic hydrolysis, immunomagnetic cell separation, fluorescence-activated cell sorting (FACS), density gradient centrifugation, immunodensity cell isolation for our customers. We choose different primary cell separation methods according to the project requirements.
- When isolating cells from intact tissues, you must first disrupt the extracellular matrix holding the cells together using proteolytic enzymes;
- Collagenase can hydrolyze collagen and is widely used for isolating cells from animal tissues;
- Hyaluronidase is often used in combination with collagenase and catalyzes hydrolysis of 1,4-β-D-glycosidic linkages;
- Elastase is used to digest tissues containing high amounts of elastin;
- Trypsin is a serine protease with a specificity for peptide bonds and is often combined with other enzymes (e.g. elastase and/or collagenase) for tissue dissociation.
- Immunomagnetic cell separation
- Due to its speed and simplicity, magnetic cell separation is one of the most commonly used methods by which scientists isolate highly purified populations of specific cell subsets;
- Targeting cells for selection or depletion using antibodies or ligands directed against specific cell surface antigens;
- Creative Proteomics uses column-based or column-free magnetic cell separation methods according to experimental needs;
- Creative Proteomics uses both positive selection and negative selection.
- Fluorescence-activated cell sorting (FACS)
- Fluorescence-activated cell sorting (FACS) is a method that uses flow cytometry and fluorescent probes to sort heterogeneous mixtures of cells;
- Sort single cells;
- Isolate cells based on intracellular markers (e.g. GFP);
- Isolate cells based on surface marker expression levels;
- Sort complex cell types with multiple markers at higher purity.
- Density gradient centrifugation
- Density gradient centrifugation relies on the varying densities of cells within a heterogeneous sample. The sample is layered on top of a density gradient medium before being centrifuged;
- Common applications include the fractionation of peripheral blood mononuclear cells, exclusion of dead cells from a cell culture, and separation of plasma from blood cells;
- Creative Proteomics offers different mart types of density gradient medium including lymphoprep, percoll, optiPrep for our customers.
- Immunodensity cell isolation
- Immunodensity cell separation, also referred to as erythrocyte rosetting, is a negative selection method that uses a combination of antibody-based labeling and density gradient centrifugation.
- Immunodensity cell separation doesn't require any specialized equipment beyond a centrifuge, can be easily incorporated into established density gradient centrifugation protocols, and can be used to isolate specific cell subsets directly from whole blood.
If you have any questions about primary cell separation methods, please contact us.
* Our services can only be used for research purposes and Not for clinical use.
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