Ficoll

Manufactured by Eurobio Scientific

Sourced in France

Ficoll is a polymer used for the separation and purification of cells, organelles, and other biological particles through density gradient centrifugation. It is a highly viscous, water-soluble substance that forms a gradient of varying density when centrifuged, allowing the desired components to be isolated based on their specific density.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Edta tube, by Merck Group (2 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Medium 199, by Thermo Fisher Scientific (2 mentions) Easysep human monocyte enrichment kit, by STEMCELL (1 mentions) Rpmi 1640 medium, by Dutscher (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Rh m csf, by Immunotools (1 mentions) Idasanutlin, by Roche (1 mentions) Topcount, by PerkinElmer (1 mentions) Anti cd3e 145 2c11, by Thermo Fisher Scientific (1 mentions) Anti cd28 antibody, by Thermo Fisher Scientific (1 mentions) Canto 2 flow cytometer, by BD (1 mentions) Propidium iodide, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Gen5 data analysis software, by Agilent Technologies (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Paxgene tube, by Qiagen (1 mentions)

Close spelling variants of this product

Ficoll density gradient centrifugation Ficoll density gradient separation Ficoll density gradient Ficoll-Paque Ficoll-Hypaque Ficoll-Hypaque density gradient Ficoll gradient Ficoll-Hypaque density gradient centrifugation Ficoll–Paque density-gradient centrifugation

21 protocols using ficoll

Isolating PBMCs from Whole Blood

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8 mL of peripheral vein whole blood were obtained from each subject, and the peripheral blood mononuclear cells (PBMCs) were isolated using density gradient centrifugation with Ficoll solution according to operating instructions (Eurobio, catalog number CMSMSL01-01). The peripheral vein whole blood was diluted with PBS (×1) in 1:1 ratio, and the mixture was transferred into a 50 mL centrifugal tube pre-filled with 8 mL Ficoll (Eurobio, catalog number CMSMSL01-01). The PBMCs were carefully collected in the middle layer of the tube following centrifugation for 30 min (500 g). Subsequently, the PBMCs were washed and centrifuged with PBS 1X for 5 min at 350 g and re-suspended in 10% DMSO / 90% foetal calf serum and cryopreserved at -80 °C. At the same time, the plasma layer was collected, centrifuged to remove cellular debris and stored at -80 °C until use. Haemolysis was quantified by assessing the presence of cell free haemoglobin by measuring the absorbance at 414 nm with a microplate spectrophotometer (Agilent BioTek Epoch with Gen5 data analysis software). Samples with signs of haemolysis (OD > 0.3) were removed from the analysis (Supplementary Material 1).

Boutabba A., Missaoui F., Dlala A., Kamoun H., Ben Salem K., Gabsi A., Rejeb H., Letessier A., Miotto B, & Marrakchi R. (2024). Circulating miR-16-5p, miR-92a-3p and miR-451a are biomarkers of lung cancer in Tunisian patients. BMC Cancer, 24, 417.

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Isolation and Differentiation of Human Monocytes

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Monocytes were purified from blood by Ficoll (Eurobio, Coutaboeuf, France) density gradient separation and by negative selection using the EasySep™ Human Monocyte Enrichment Kit (Stem Cell, Grenoble, France). Monocytes were suspended in RPMI 1640 medium (Dutscher, Brumate, France) supplemented with 10% heat-inactivated fetal calf serum (FCS; Dutscher, Brumate, France), 1% l-glutamine (Life Technologies, Carlsbad, CA, USA), and 0.2 μg/mL of recombinant human macrophage colony stimulating factor (rh-M-CSF, Immunotools, Friesoythe, Germany). Cells were seeded into 48-well culture plates at a density of 2.5 × 10⁵ and were incubated at 37 °C in a humidified 5% CO₂ atmosphere for six days.

Buisson A., Douadi C., Ouchchane L., Goutte M., Hugot J.P., Dubois A., Minet-Quinard R., Bouvier D., Bommelaer G., Vazeille E, & Barnich N. (2019). Macrophages Inability to Mediate Adherent-Invasive E. coli Replication is Linked to Autophagy in Crohn’s Disease Patients. Cells, 8(11), 1394.

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Monocyte Isolation from Peripheral Blood

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Peripheral blood was obtained from healthy adults and from patients with Crohn's disease, peripheral blood mononuclear cells (PBMC) were isolated by density gradient centrifugation using Ficoll (Eurobio). Blood monocytes were isolated from PBMC by MACS sorting with PE anti-CD14 (rmC5-3) (Miltenyi). The purity of enriched monocytes samples were evaluated by flow cytometry.

Petit V., Parcelier A., Mathé C., Barroca V., Torres C., Lewandowski D., Ferri F., Gallouët A.S., Dalloz M., Dinet O., Boschetti G., Vozenin M.C, & Roméo P.H. (2019). TRIM33 deficiency in monocytes and macrophages impairs resolution of colonic inflammation. EBioMedicine, 44, 60-70.

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Expansion of CD34+ MPN Cells with Idasanutlin

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CD34⁺ cells from MPN patients after NGS analysis were prospectively cryopreserved. Peripheral mononuclear cells were isolated from whole blood using Ficoll (Eurobio) and CD34⁺cells sorted using a column-free immunomagnetic approach (EasySep, StemCell). 10⁵ CD34⁺ cells were cultured in CTS^TM StemPro^TM hematopoietic stem cell expansion medium (Thermo Fisher) in which the following cytokines were added: IL-3 (50 ng/mL), stem cell factor (100 ng/mL), thrombopoietin (20 ng/mL) IL-6 (50 ng/mL), and FLT-3 ligand (100 ng/mL). Idasanutlin (provided by Roche) was added at 20 nM. After 10 days of culture, cells were harvested, DNA extracted, and submitted to NGS analysis. This test is developed under the patent application EP 21 305 499.2.

Maslah N., Verger E., Giraudier S., Chea M., Hoffman R., Mascarenhas J., Cassinat B, & Kiladjian J.J. (2022). Single-cell analysis reveals selection of TP53-mutated clones after MDM2 inhibition. Blood Advances, 6(9), 2813-2823.

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T Cell Proliferation Assay with Activation

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Peripheral blood mononuclear cells (PBMC) were isolated by centrifugation using Ficoll (Eurobio) density gradient. The quantification of T cells was obtained by staining PBMC with a CD3e (145-2C11) antibody (Becton Dickinson). A 96-well round-bottom plate was precoated with an anti-CD3e (145-2C11, 5 µg/mL, eBioscience) for 2 h at 37 °C. T cells (2.10⁴ cells/well) were added and incubated for 48 h with an anti-CD28 antibody (37.51, 2.5 µg/mL, eBioscience) or complete media for control well in a humidified environment with 5% CO₂ at 37 °C. Then, [3H] thymidine was added in the culture medium (1 µCi/well, PerkinElmer) 16 h before harvesting cells on a fiberglass filter using an automated cell harvester (PerkinElmer). Incorporated radioactivity was measured in a direct beta counter (Top Count, PerkinElmer). The results were expressed as a proliferation ratio between counts per minute measured in stimulated and nonstimulated wells for the same sample.

Reizine F., Grégoire M., Lesouhaitier M., Coirier V., Gauthier J., Delaloy C., Dessauge E., Creusat F., Uhel F., Gacouin A., Dessauge F., Le Naoures C., Moreau C., Bendavid C., Daniel Y., Petitjean K., Bordeau V., Lamaison C., Piau C., Cattoir V., Roussel M., Fromenty B., Michelet C., Le Tulzo Y., Zmijewski J., Thibault R., Cogné M., Tarte K, & Tadié J.M. (2022). Beneficial effects of citrulline enteral administration on sepsis-induced T cell mitochondrial dysfunction. Proceedings of the National Academy of Sciences of the United States of America, 119(8), e2115139119.

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PBMCs Cytotoxicity Assay with Pig LIS1 Antibodies

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hPBMCs were isolated from 3 healthy donors by Ficoll (Eurobio, France) density gradient centrifugation. 2.5 × 10⁵ PBMC were incubated with increasing doses of purified pig LIS1 IgG antibodies or control nonimmune (67p) IgG in phosphate buffer solution (PBS) 1% bovine serum albumin (BSA) (Merck, France) for 30 minutes at 4°C. PBMC were then washed twice with PBS 1% BSA and resuspended on ice in 25 µL of neat human, mouse or rat serum. After 30 minutes incubation at 37°C, PBMC were washed twice with PBS 1% BSA on ice and resuspended in 100 µL of propidium iodide at 4 µg/mL (Merck, France). Cell cytotoxicity was determined by the % of PI-positive cells on a Canto II flow cytometer (BD Biosciences).

Ménoret S., Ouisse L.H., Tesson L., Remy S., Usal C., Guiffes A., Chenouard V., Royer P.J., Evanno G., Vanhove B., Piaggio E, & Anegon I. (2020). In Vivo Analysis of Human Immune Responses in Immunodeficient Rats. Transplantation, 104(4), 715-723.

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Whole Blood and PBMC RNA Extraction

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RNAs were extracted from whole blood (PAXgene tubes, Qiagen, Courtaboeuf, France) or PBMCs as previously isolated from EDTA tubes (Sigma Aldrich, Saint-Quentin Fallavier, France) using Ficoll (Eurobio, Lees Ulis, France) gradient and centrifugation as previously described [18 (link)] with RNeasy Mini Kit with a DNase I treatment to eliminate DNA contaminants and according to the manufacturer instructions (Qiagen, Courtaboeuf, France).

Melenotte C., Mezouar S., Ben Amara A., Benatti S., Chiaroni J., Devaux C., Costello R., Kroemer G., Mege J.L, & Raoult D. (2019). A transcriptional signature associated with non-Hodgkin lymphoma in the blood of patients with Q fever. PLoS ONE, 14(6), e0217542.

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PBMC Isolation and Monocyte Purification

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Peripheral blood mononuclear cells (PBMCs) were recovered from blood samples using Ficoll (Eurobio, Les Ulis, France) density gradient centrifugation followed by harvesting of PBMCs. Monocytes were isolated by adherence to culture dishes for 2 hours at 37°C and directly lysed for qRT-PCR without being harvested.

Osman I.O., Melenotte C., Brouqui P., Million M., Lagier J.C., Parola P., Stein A., La Scola B., Meddeb L., Mege J.L., Raoult D, & Devaux C.A. (2021). Expression of ACE2, Soluble ACE2, Angiotensin I, Angiotensin II and Angiotensin-(1-7) Is Modulated in COVID-19 Patients. Frontiers in Immunology, 12, 625732.

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Isolation and Cryopreservation of PBMCs

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Peripheral blood samples were collected on lithium heparin. PBMCs were isolated by density gradient centrifugation (2,200 rpm without break for 30 minutes) using Ficoll (Eurobio Scientific, Les Ulis, France). After centrifugation, cells were washed with Phosphate-buffered saline (PBS) (Thermo Fisher scientific, Illkirch, France). The pellet was resuspended in PBS and cells were centrifuged at 1,900 rpm for 5 minutes. Finally, the PBMCs pellet was frozen in a medium containing 90% of Fetal Bovine Serum (FBS) (GIBCO, Thermo Fisher scientific, Illkirch, France) and 10% of dimethyl sulfoxide (DMSO) (Sigma Aldrich, St. Quentin Fallavier, France).

de Cevins C., Luka M., Smith N., Meynier S., Magérus A., Carbone F., García-Paredes V., Barnabei L., Batignes M., Boullé A., Stolzenberg M.C., Pérot B.P., Charbit B., Fali T., Pirabakaran V., Sorin B., Riller Q., Abdessalem G., Beretta M., Grzelak L., Goncalves P., Di Santo J.P., Mouquet H., Schwartz O., Zarhrate M., Parisot M., Bole-Feysot C., Masson C., Cagnard N., Corneau A., Brunaud C., Zhang S.Y., Casanova J.L., Bader-Meunier B., Haroche J., Melki I., Lorrot M., Oualha M., Moulin F., Bonnet D., Belhadjer Z., Leruez M., Allali S., Gras-Leguen C., de Pontual L., Fischer A., Duffy D., Rieux-Laucat F., Toubiana J, & Ménager M.M. (2021). A monocyte/dendritic cell molecular signature of SARS-CoV-2-related multisystem inflammatory syndrome in children with severe myocarditis. Med (New York, N.y.), 2(9), 1072-1092.e7.

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Isolation of Human Monocytes

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Blood was obtained from healthy donors from the Etablissement Français du Sang Auvergne Rhône Alpes, France, under the convention EFS 16–2066. Peripheral blood mononuclear cells were isolated from 450 ml of freshly drawn venous human peripheral blood by Ficoll (Eurobio) density gradient centrifugation. Cells were washed three times in sterile PBS (Thermo Fisher Scientific) and resuspended in separation medium (PBS, pH 7.2, supplemented with 0.5% BSA and 2 mM EDTA). Monocyte purification was performed by CD14-positive selection using a human CD14 microbeads purification kit (Miltenyi Biotec) following the manufacturer’s recommendations. Cell suspensions were always >95% CD14⁺ as determined by flow-cytometric analysis.

Omarjee O., Mathieu A.L., Quiniou G., Moreews M., Ainouze M., Frachette C., Melki I., Dumaine C., Gerfaud-Valentin M., Duquesne A., Kallinich T., Tahir Turanli E., Malcus C., Viel S., Pescarmona R., Georgin-Lavialle S., Jamilloux Y., Larbre J.P., Sarrabay G., Magnotti F., Rice G.I., Bleicher F., Reboulet J., Merabet S., Henry T., Crow Y.J., Faure M., Walzer T, & Belot A. (2021). LACC1 deficiency links juvenile arthritis with autophagy and metabolism in macrophages. The Journal of Experimental Medicine, 218(3), e20201006.

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