Ficoll density gradient

Manufactured by Harvard Bioscience

Sourced in Germany

Ficoll density gradient is a laboratory technique used for the separation and purification of cells, proteins, and other biomolecules based on their density. It utilizes a water-soluble, synthetic polymer called Ficoll to create a density gradient medium, enabling the separation of different components within a sample.

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Lab products found in correlation

Fetal calf serum (fcs), by GE Healthcare (1 mentions) Dimethyl sulfoxide (dmso), by Carl Roth (1 mentions) Ethylene diamine tetraacetic acid (edta), by GE Healthcare (1 mentions) Nh4 heparin monovette tubes, by Sarstedt (1 mentions) Fetal calf serum (fcs), by Carl Roth (1 mentions) Spin column, by Qiagen (1 mentions) One lambda, by Thermo Fisher Scientific (1 mentions) 24 well flat bottom tissue culture plates, by Sarstedt (1 mentions) Heat inactivated fetal calf serum, by Harvard Bioscience (1 mentions) Cd16 microbeads, by Miltenyi Biotec (1 mentions) Penicillin, by Harvard Bioscience (1 mentions) Streptomycin, by Harvard Bioscience (1 mentions) L glutamine, by Harvard Bioscience (1 mentions)

Spelling variants of this product

Ficoll (64 citations) Ficoll gradient (11 citations) Ficoll density gradient centrifugation (11 citations) Ficoll-Hypaque density gradient centrifugation (5 citations) Ficoll-Hypaque density gradient (4 citations) Ficoll-Paque density gradient (3 citations)

4 protocols using ficoll density gradient

PBMC Isolation from CRC Samples

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Blood samples from CRC patients or healthy donors were collected in NH₄-Heparin Monovette tubes (Sarstedt). PBMCs were isolated by Ficoll density gradient (Biochrom AG) centrifugation. Isolated cells were washed with PBS containing 2 mmol/l EDTA and 2% FCS (PAA Laboratories), and cryopreserved in cell culture medium containing 10% FCS and 10% DMSO (Carl Roth GmbH) until further analysis.
Patients and healthy donors gave written informed agreement and analyses of the samples were approved by the Cantonal Ethics Committee of Bern (2017-01821) or the Ethics Committee of the Medical Faculty of the University Duisburg Essen (AZ 05-2882).

Pastille E., Wasmer M.H., Adamczyk A., Vu V.P., Mager L.F., Phuong N.N., Palmieri V., Simillion C., Hansen W., Kasper S., Schuler M., Muggli B., McCoy K.D., Buer J., Zlobec I., Westendorf A.M, & Krebs P. (2019). The IL-33/ST2 pathway shapes the regulatory T cell phenotype to promote intestinal cancer. Mucosal Immunology, 12(4), 990-1003.

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HLA-typing of HBV Patients

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In a multi-center effort for analysis of HLA class I-associated selection pressure 464 HBV patients were recruited at the Hepatology Units in Düsseldorf, Essen, Hamburg, Hannover, Freiburg (all Germany) and London (UK); (Supplemental Table 1). Only HBV genotype A and D infected patients were included. Informed consent was obtained from each patient, and the study protocol was approved by the local ethics committee of the Medical Faculty of Düsseldorf in accordance with the guidelines of the Declaration of Helsinki. Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll density gradient centrifugation (Biocoll; Biochrom) (9 (link)). DNA for HLA-typing was extracted from PBMCs using spin columns (Qiagen). High resolution (second field) HLA-A and HLA-B typing was performed by use of sequence-specific oligonucleotides (LABType™) methodology (19 (link)), provided by One Lambda (Thermo Fisher Inc.) at department of transfusion medicine of the University Hospital Essen.

Walker A., Schwarz T., Brinkmann-Paulukat J., Wisskirchen K., Menne C., Alizei E.S., Kefalakes H., Theissen M., Hoffmann D., Schulze zur Wiesch J., Maini M.K., Cornberg M., Kraft A.R., Keitel V., Bock H.H., Horn P.A., Thimme R., Wedemeyer H., Heinemann F.M., Luedde T., Neumann-Haefelin C., Protzer U, & Timm J. (2022). Immune escape pathways from the HBV core18-27 CD8 T cell response are driven by individual HLA class I alleles. Frontiers in Immunology, 13, 1045498.

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Isolation of Peripheral Blood Mononuclear Cells

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Peripheral blood mononuclear cells were isolated from buffy coats derived from blood donation bottles of six healthy volunteers by density centrifugation on Ficoll density gradient (Biochrom AG, Berlin, Germany) as previously described (Spiliopoulou et al., 2012 (link)). Briefly, collected mononuclear cells were washed in PBS and resuspended in RPMI-1640 medium supplemented with 10% heat-inactivated fetal calf serum (Biochrom AG) and 2 mM L-glutamine (HyClone), (CM). Cells at a density of 1 × 10⁶ cells/mL per well, were then seeded in 24-well flat bottom tissue culture plates (Sarstedt, Nümbrecht, Germany) and cultured at 37°C in a humidified, 5% CO₂ atmosphere.

Kolonitsiou F., Papadimitriou-Olivgeris M., Spiliopoulou A., Drougka E., Jelastopulu E., Anastassiou E.D, & Spiliopoulou I. (2019). Methicillin-Resistant Staphylococcus aureus ST80 Induce Lower Cytokine Production by Monocytes as Compared to Other Sequence Types. Frontiers in Microbiology, 9, 3310.

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Isolation of Peripheral Blood Eosinophils

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Peripheral blood eosinophils were obtained from heparin-anticoagulated venous blood and prepared following the protocol of Raap et al. (20) . Briefly, blood was layered on a Ficoll density gradient (Biochrom, Berlin, Germany). After discarding the supernatant, pellets were resuspended in lysis buffer. Eosinophils were isolated from the remaining cells by a negative immunomagnetic bead selection with CD16 microbeads (Miltenyi Biotech, Bergisch Gladbach, Germany), following the manufacturer's protocol. The purity of the eosinophils was > 98%, as assessed by fluorescenceactivated cell sorting (FACS) analysis (CD66b/CD16) and Kimura staining (21) . The viability was greater than 99%, as assessed by trypan blue dye exclusion. If not otherwise described, 2 × 10 5 eosinophils/200 µl were seeded into wells of 96 well plates and cultivated in RPMI 1640 with 10% heat inactivated foetal calf serum (FCS) including 2 mM L-glutamine, 10,000 U/ml penicillin, and 10 mg/ml streptomycin (all Seromed, Biochrom, Berlin, Germany) at 37°C and 5% CO 2 .

Engmann J., Rüdrich U., Behrens G., Papakonstantinou E., Gehring M., Kapp A, & Raap U. (2017). Increased Activity and Apoptosis of Eosinophils in Blister Fluids, Skin and Peripheral Blood of Patients with Bullous Pemphigoid. Acta dermato-venereologica, 97(4).

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