Ficoll paque plus

Manufactured by STEMCELL

Sourced in Canada

Ficoll-Paque Plus is a sterile, pyrogen-tested density gradient medium used for the isolation of mononuclear cells from whole blood, bone marrow, or other tissues. It facilitates the separation of different cell populations based on their density differences.

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Lab products found in correlation

Vacutainer tube, by BD (3 mentions) Ficoll paque plus, by GE Healthcare (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) Penicillin, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Human pan dc pre enrichment kit, by STEMCELL (2 mentions) Cd34 microbead kit, by Miltenyi Biotec (2 mentions) Magnetic bead, by Miltenyi Biotec (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Rosettesep human b cell enrichment cocktail, by STEMCELL (1 mentions) Insulin transferrin sodium selenite supplement, by Merck Group (1 mentions) Facsaria, by BD (1 mentions) Rhtslp, by R&D Systems (1 mentions) Sigmafast protease inhibitor cocktail, by Merck Group (1 mentions) Mouse anti β actin, by Merck Group (1 mentions) Rabbit anti acetylated histone h3, by Merck Group (1 mentions) Human cd34 cd38 isolation kit, by Miltenyi Biotec (1 mentions) Anti human cd34 antibody, by Miltenyi Biotec (1 mentions) Magnetic activated cell sorting (macs), by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

Ficoll-Paque (27 citations) Ficoll (25 citations)

23 protocols using ficoll paque plus

Isolating PBMCs from Blood

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Blood samples were collected from the subjects in standard 10-mL heparin-treated vacutainer tubes (Vacutainer® tubes; Becton Dickinson, Stockholm, Sweden). Peripheral blood mononuclear cells (PBMCs) were isolated from each sample using Ficoll-Paque density gradient centrifugation (Ficoll-Paque™ plus; Stem Cell Technologies, Vancouver, BC, Canada).

Park S.J., Park M.J., Park S., Lee E.S, & Lee D.Y. (2023). Integrative metabolomics of plasma and PBMCs identifies distinctive metabolic signatures in Behçet’s disease. Arthritis Research & Therapy, 25, 5.

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Timing and Processing of CTC Samples

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Pre-treatment, baseline blood specimens (Pre-RT) for CTC analysis were collected within 1 week before starting RT, typically on the day of CT simulation for RT planning or on the day of pre-treatment patient set-up. During RT, specimens were collected at up to 3 time points, including during the first week of RT (1W-RT), mid-way through RT (Mid-RT), and during the last week of RT (End-RT). Post-treatment (Post-RT) blood specimens were drawn 4 to 12 weeks after the completion of RT during follow-up clinic appointments.
Approximately 12 mL of whole peripheral blood, drawn from either healthy donors or cancer patients, was collected into heparin-treated BD Vacutainer tubes to prevent coagulation, except for the first patient enrolled, whose baseline specimen was collected into EDTA-treated BD Vacutainer tubes. Blood specimens were kept at ambient temperature, and shipped from UNC to UIC via overnight express to analyze the specimens within 24 hours after blood collection. Mononuclear cells including CTCs in buffy coat were separated from whole blood using Ficoll-Paque Plus (Stemcell Technologies Inc., Vancouver, Canada) as described previously (17 (link)). After washing the buffy coat twice with 2% FBS-containing PBS, the recovered cells were suspended in 0.2 mL of the complete DMEM media and used for subsequent experiments.

Myung J.H., Eblan M.J., Caster J.M., Park S.J., Poellmann M.J., Wang K., Tam K.A., Miller S.M., Shen C., Chen R.C., Zhang T., Tepper J.E., Chera B.S., Wang A.Z, & Hong S. (2018). Multivalent binding and biomimetic cell rolling improves the sensitivity and specificity of circulating tumor cell capture. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(11), 2539-2547.

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Pediatric Leukemia Cell Isolation Protocol

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Bone marrow or peripheral blood samples from untreated children initially diagnosed with BCP-ALL, T-ALL or AML were collected from the Czech Pediatric Hematology Centers. The inclusion criteria were the percentage of blasts higher than 80% and high cellularity. Within 24 h after aspiration, without freezing, the mononuclear cells were isolated by density gradient centrifugation using Ficoll-Paque PLUS (GE Healthcare, UK). All samples were obtained with the informed consent of the children’s parents or guardians. The study no. 201528848A was approved by the Ethical Committee of the University Hospital Motol, Prague, Czech Republic. Healthy controls were isolated from buffy coats (mixture of healthy individuals) using Ficoll-Paque PLUS (GE Healthcare, UK). To enrich samples for B-lymphocytes, buffy coat was pre-treated with RosetteSep™ Human B Cell Enrichment Cocktail (StemCell Technologies, USA) prior to Ficoll-Paque PLUS.
The isolated blasts were maintained in RPMI-1640 medium with GlutaMAX™ supplemented with 10% fetal calf serum, penicillin (100 U/mL) and streptomycin (100 μg/mL). For the MTS assay, insulin-transferrin-sodium selenite supplement was added to the culture media (Sigma-Aldrich, St Louis, MO, USA).

Hlozkova K., Pecinova A., Alquezar-Artieda N., Pajuelo-Reguera D., Simcikova M., Hovorkova L., Rejlova K., Zaliova M., Mracek T., Kolenova A., Stary J., Trka J, & Starkova J. (2020). Metabolic profile of leukemia cells influences treatment efficacy of L-asparaginase. BMC Cancer, 20, 526.

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Isolation of Tumor Cells from Whole Blood

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The heparin-treated
blood was kept at 4 °C in a refrigerator and the experiments
were performed within 48 h after drawing. Fluorescence-labeled cancer
cells were spiked to 3 mL of whole blood as a final concentration
of 1 × 10⁵ tumor cells/mL blood. Mononuclear cells
including tumor cells in buffy coat was separated from whole blood
using Ficoll-Paque Plus (Stemcell Technologies Inc., Vancouver, Canada)
as described in section 7 of Supporting Information. Briefly, the blood samples loaded with Ficoll for separation were
centrifuged at 20 °C for 20 min at 1,500× g with brake function
off. After the buffy coat was washed twice with the FBS/heparin-included
PBS buffer via centrifuge, the recovered cells were suspended with
3 mL of the complete cell culture media and used for subsequent experiments.
Studies using human blood were reviewed and approved by UIC institutional
review board (IRB) (protocol 2012-0139).

Myung J.H., Gajjar K.A., Chen J., Molokie R.E, & Hong S. (2014). Differential Detection of Tumor Cells Using a Combination of Cell Rolling, Multivalent Binding, and Multiple Antibodies. Analytical Chemistry, 86(12), 6088-6094.

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Isolation and Analysis of Circulating Tumor Cells

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Whole blood drawn from healthy donors was collected in heparin-treated tubes and kept at ambient temperature. Studies using human blood were reviewed and approved by UIC institutional review board (IRB) (protocols #2012–0139 for tumor cell-spiked blood specimens and #2013–1033 for clinical PCa patient blood specimens). Fluorescence-labeled BCa or PCa cells were spiked into 3 mL of whole blood at a final concentration of 1 × 10⁵ tumor cells/mL blood. Mononuclear cells including tumor cells in buffy coat were separated from whole blood using Ficoll-Paque Plus (Stemcell Technologies Inc., Vancouver, Canada) as described in our previous publication.^{21 (link)} After washing the buffy coat twice with the 2% FBS-containing PBS, the recovered cells were suspended in 3 mL of the complete cell culture media and used for subsequent experiments. For the clinical samples, 12 mL of the blood specimen obtained from prostate cancer patients were used for separation and resuspended with 12 mL of complete media. The experiments were performed within 48 hrs after blood drawing.

Myung J.H., Cha A., Tam K.A., Poellmann M., Borgeat A., Sharifi R., Molokie R.E., Votta-Velis G, & Hong S. (2019). A Dendrimer-based Platform for Effective Capture of Tumor Cells after TGFβ1-induced Epithelial-Mesenchymal Transition. Analytical chemistry, 91(13), 8374-8382.

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Isolation and Culture of Human Dendritic Cells

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DC were enriched from PBMC obtained after Ficoll-Paque Plus density gradient centrifugation (Stemcell Technologies; Vancouver, BC, Canada) by negative selection with monoclonal antibodies (mAb) to CD3, CD9, CD14, CD16, CD19, CD34, CD56, CD66b, and glycophorin A (Human pan-DC pre-enrichment kit; Stemcell Technologies). Cells were labeled with anti-human lineage cocktail-FITC (CD3, CD14, CD16, CD19, CD20 and CD56); CD123-PE (9F5) and CD11c-APC (S-HCL-3) (BD Biosciences; Franklin Lakes, NJ), and HLA-DR-APC-eflour780 (LN3) (Sigma-Aldrich; St. Louis, MO); lin⁻CD123⁻HLA-DR⁺CD11c⁺ DC were sorted with FACS Aria (BD Bioscience). DC were seeded at 100 × 10³ cells/well in 200 μl of RPMI with 10% human AB serum, and cultured with medium alone or in the presence of 20 ng/ml of rhTSLP (R&D), or tumor-derived products. After 48 hours DC were harvested, washed, and analyzed or used in experiments.

Wu T.C., Xu K., Banchereau R., Marches F., Yu C.I., Martinek J., Anguiano E., Pedroza-Gonzalez A., Snipes G.J., O’Shaughnessy J., Nishimura S., Liu Y.J., Pascual V., Banchereau J., Oh S, & Palucka K. (2014). Reprogramming tumor-infiltrating dendritic cells for CD103+CD8+ mucosal T cell differentiation and breast cancer rejection. Cancer immunology research, 2(5), 487-500.

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Isolation of PBMCs and DCs

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PBMCs were isolated from leukapheresis products using Ficoll-Paque Plus density gradient centrifugation (Stemcell Technologies, Vancouver, BC, Canada). DCs were first enriched using human Pan-DC Pre-Enrichment Kit (StemCell Technologies) before staining for FACS-sorting.

Yu C.I., Becker C., Metang P., Marches F., Wang Y., Toshiyuki H., Banchereau J., Merad M, & Palucka K. (2014). Human CD141+ DCs induce CD4+ T cells to produce type 2 cytokines. Journal of immunology (Baltimore, Md. : 1950), 193(9), 4335-4343.

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Isolation and Analysis of Histone Acetylation

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Blood samples for MC fractions were collected in EDTA-coated BD Vacutainer tubes. Diluted blood was fractionated using Ficoll-Paque PLUS (Stemcell Technologies cat #07967), according to the manufacturer’s instructions and using a single wash step. MC pellets were washed in 1X SigmaFast protease inhibitor cocktail (Sigma cat #S8820), and stored at −80°C. Protein lysates were prepared from these pellets in RIPA buffer with 1X protease inhibitor. Histone acetylation and levels of the control beta-actin were assayed by SDS-PAGE and Western blotting (rabbit anti-acetylated histone H3, Millipore cat #06-599, mouse anti-beta-actin, Sigma cat #A5441).

Alumkal J.J., Slottke R., Schwartzman J., Cherala G., Munar M., Graff J.N., Beer T.M., Ryan C.W., Koop D.R., Gibbs A., Gao L., Flamiatos J.F., Tucker E., Kleinschmidt R, & Mori M. (2014). A phase II study of sulforaphane-rich broccoli sprout extracts in men with recurrent prostate cancer. Investigational new drugs, 33(2), 480-489.

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Isolation and Purification of CD34+ Cells

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Bone marrow samples were collected from patients newly diagnosed with CML as part of an institutionally approved protocol for cell sample collection (Centre Hospitalier Nice, 06204 Nice, France). Cord blood and bone marrow samples were collected from healthy donors with informed consent. Mononuclear cells were isolated by density centrifugation (Ficoll-Paque Plus, STEMCELL Technologies), washed with PBS with 5% fetal calf serum (FCS). For clonogenic assays CD34⁺ cells were positively enriched with CD34 microbeads (CD34 MicroBead Kit, Miltenyi Biotec). For LTC-IC, CD34⁺ cells were first positively enriched and then depleted for CD38⁺ cells (human CD34⁺CD38⁻ isolation kit, Miltenyi Biotec). The CD34⁺ cells, after sorting, were stained with anti-human-CD34 antibody (Miltenyi Biotec, 130-113-741) and checked by flow cytometry to ensure purity (always above 96%).

Muselli F., Mourgues L., Morcos R., Rochet N., Nebout M., Guerci-Bresler A., Faller D.V., William R.M., Mhaidly R., Verhoeyen E., Legros L., Peyron J.F, & Mary D. (2021). Combination of PKCδ Inhibition with Conventional TKI Treatment to Target CML Models. Cancers, 13(7), 1693.

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Isolation of PBMC Subsets

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Peripheral blood mononuclear cells (PBMC) were obtained by Ficoll-Paque™ Plus (GE Life Sciences) centrifugation of the blood samples. Monocytes (CD14⁺ cells) were isolated by MACS (Miltenyi); NK cells and T cells were isolated using RosetteSep (StemCell) and Ficoll-Paque™ Plus centrifugation. Purity of isolated cells was always above 90%, as assessed by flow cytometry as CD14⁺ cells (monocytes), CD3^-CD56⁺ cells (NK cells) or CD3⁺ cells (T cells).

Sierra J.M., Secchiari F., Nuñez S.Y., Iraolagoitia X.L., Ziblat A., Friedrich A.D., Regge M.V., Santilli M.C., Torres N.I., Gantov M., Trotta A., Ameri C., Vitagliano G., Pita H.R., Rico L., Rovegno A., Richards N., Domaica C.I., Zwirner N.W, & Fuertes M.B. (2021). Tumor-Experienced Human NK Cells Express High Levels of PD-L1 and Inhibit CD8+ T Cell Proliferation. Frontiers in Immunology, 12, 745939.

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