Ficoll paque plus

Manufactured by Greiner

Sourced in Germany

Ficoll-Paque Plus® is a density gradient medium used for the separation and isolation of cells, subcellular organelles, and macromolecules. It is a sterile, pyrogen-free solution composed of sucrose and Ficoll polymer.

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Lab products found in correlation

Leucosep tube, by Greiner (2 mentions) Vp h100k, by Terumo (1 mentions) Aim 5 medium, by Thermo Fisher Scientific (1 mentions) Rpmi media, by Merck Group (1 mentions) Rbc lysis solution, by Miltenyi Biotec (1 mentions)

5 protocols using ficoll paque plus

Toxoplasma Infection Analysis in Adults

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Clinical and serological analyses were performed on a group of 43 women and 21 men in the age range of 20–67 years (mean value 31.6 years; median 28). Only those patients who voluntarily signed a written agreement were involved in the project. The study was approved by the Local Committee of Ethics in Science (licence number 11 ⁄ 10 ⁄ 2005) and conducted in accordance with the Helsinki Declaration.
Clinical specimens for the study were collected from each patient and consisted of serum samples (used to determine the presence of the anti-Toxoplasma IgG antibody) and peripheral blood samples stabilized with EDTA (as a source of peripheral blood mononuclear cells – PBMCs). PBMCs were isolated from the blood samples using a Leucosept-Tube with Ficoll-Paque Plus® (Greiner Bio-One, Germany) according to the manufacturer’s instructions. The final number of PBMCs was adjusted to 2.5 × 10⁶ cells ⁄mL and they were used as host cells for T. gondii in vitro proliferation tests.

Dzitko K., Grzybowski M.M., Pawełczyk J., Dziadek B., Gatkowska J., Stączek P, & Długońska H. (2015). Phytoecdysteroids as modulators of the Toxoplasma gondii growth rate in human and mouse cells. Parasites & Vectors, 8, 422.

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PBMC Isolation and Plasma Separation from Heparinized Blood

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Using Leucosep tubes pre-filled with Ficoll-Paque Plus (Greiner; 163288), PBMCs and plasma were separated from blood samples collected in heparin-coated tubes (TERUMO; VP-H100K) as previously described⁴⁸. Briefly, 25 mL of blood and 12 mL of AIM-V medium (Thermo; 12055091) were mixed, added to Leucosep tubes, and centrifuged at 1000 × g at room temperature for 10 min. The upper yellowish plasma solution was collected and stored at –20 °C. The white layer containing PBMCs were washed twice with 22 mL of AIM-V medium by 7 min centrifugation, once at 600 × g and a second time at 400×g, and then cells were resuspended in 500 μL of CTL test medium (Cellular Technology Limited (CTL); CTLT-010). Fresh PBMCs were used for IFN-γ ELISpot assays, and the remaining PBMCs were stored in liquid nitrogen until use for other assays.

Hirota M., Tamai M., Yukawa S., Taira N., Matthews M.M., Toma T., Seto Y., Yoshida M., Toguchi S., Miyagi M., Mori T., Tomori H., Tamai O., Kina M., Sakihara E., Yamashiro C., Miyagi M., Tamaki K., Wolf M., Collins M.K., Kitano H, & Ishikawa H. (2023). Human immune and gut microbial parameters associated with inter-individual variations in COVID-19 mRNA vaccine-induced immunity. Communications Biology, 6, 368.

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PBMC Isolation from Heparinized Blood

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Twenty milliliters (ml) of heparinized venous blood were collected, briefly centrifuged, plasma separated then packed cells processed for PBMCs isolation. Each aliquot of packed cells was diluted with equal volume of Roswell Park Memorial Institute (RPMI) media (Sigma), layered over Ficoll-Paque plus on Leucosep tubes (Greiner), and PBMCs separated by density gradient centrifugation. Cells in the supernatant were harvested and washed three times with phosphate-buffered saline. Finally, cells were resuspended in 1 ml RPMI and manually counted on a hemocytometer after staining with trypan blue to check viability of the cells. Dead cells appeared blue as the trypan blue passed through the damaged cell membrane while viable cells were colorless. The average viability of cells was 98%.

Tamene W., Wassie L., Marconi V.C., Abebe M., Kebede A., Sack U, & Howe R. (2024). Protein Expression of TLR2, TLR4, and TLR9 on Monocytes in TB, HIV, and TB/HIV. Journal of Immunology Research, 2024, 9399524.

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Isolation of PBMCs from Human Blood Samples

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Human peripheral blood samples were collected from anonymous healthy donors and patients with MG at Keio University Hospital (Figure 1A). PBMCs were separated using a LeucoSep tube filled with Ficoll-Paque-plus (Greiner Bio-One, Kremsmünster, Austria), according to the manufacturer’s instructions. Briefly, blood samples were transferred into tubes and centrifuged at 1000 ×g for 10 min at room temperature. PBMCs from the interfacial layer were collected, washed with 1% bovine serum albumin (BSA)/phosphate-buffered saline (PBS), resuspended in 1% BSA/PBS, and filtered through 40-μm FLOWMI Cell Strainers (Bel-Art Products, South Wayne, NJ, USA). scRNA-seq analysis was performed on cells resuspended in 1% BSA/PBS at a concentration of 1 × 10⁶ cells/ml. PBMCs contaminated with red blood cells (RBCs) were subjected to RBC lysis using RBC lysis solution (Miltenyi Biotec, Bergisch Gladbacblooh, Germany). RBCs were removed by incubating PBMCs in 10 volumes of 10X RBC lysis solution diluted with double-distilled H₂O for 10 min at room temperature. To remove the lysis buffer, PBMCs were washed with 1% BSA/PBS. Heparin plasma was isolated from the blood of the same patient in a different tube than that used for PBMC collection through centrifugation at 3000 rpm for 10 min at 4°C.

Okuzono Y., Miyakawa S., Itou T., Sagara M., Iwata M., Ishizuchi K., Sekiguchi K., Motegi H., Oyama M., Warude D., Kikukawa Y, & Suzuki S. (2024). B-cell immune dysregulation with low soluble CD22 levels in refractory seronegative myasthenia gravis. Frontiers in Immunology, 15, 1382320.

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Chronic Alopecia Areata PBMC Analysis

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Peripheral blood mononuclear cells (PBMCs) were taken from 11 non-atopic patients with chronic-phase AA (hair loss for longer than 6 months and Severity of Alopecia Tool (SALT) score > 25), 13 patients with chronic-phase AA and extrinsic AD, 6 patients with chronic-phase AA and intrinsic AD, and 9 healthy volunteers.
PBMCs were isolated from heparinized venous blood by centrifugation with Ficoll-Paque PLUS in LeucoSep tubes (Greiner Bio-One, Frickenhausen, Germany). Skin samples were obtained from one AA patient with extrinsic AD.

Kageyama R., Ito T., Hanai S., Morishita N., Nakazawa S., Fujiyama T., Honda T, & Tokura Y. (2021). Immunological Properties of Atopic Dermatitis-Associated Alopecia Areata. International Journal of Molecular Sciences, 22(5), 2618.

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