Separating Solutions   Pancoll In many cases the isolation of cells is the first step for gene expression studies or in diagnostic procedures. Besides biological separation techniques physical separation methods are most commonly used. These methods use physical differences such as size and weight of the particles to be separated. For this purpose so-called separating solutions (= centrifugation media) are used. Our Pancoll separating solutions contain a polysaccharide with a molecular weight of 400,000 daltons; this hydrophilic polymer allows for production of aqueous solutions for cell separation with a density of up to 1.2 g/ml. PAN-Biotech offers a variety of ready-to-use products with a density of 1.063 g/ml up to 1.091 g/ml for a very wide range of cell separation applications.     Description Size Product number Datasheet Pancoll human, density 1.077 g/ml 100 ml500 ml P04-60100P04-60500 Pancoll human for granulocytes, density 1.119 g/ml 100 ml500 ml P04-60110P04-60150 Pancoll mouse, density 1.086 g/ml 100 ml500 ml P04-64100P04-64500 Pancoll rat, density 1.091 g/ml 100 ml500 ml P04-65100P04-65500 Pancoll animal, density 1.077 g/ml 100 ml500 ml P04-63100P04-63500 Pancoll monocytes, density 1.068 g/ml 100 ml500 ml P04-68100P04-68500 Pancoll platelets, density 1.063 g/ml 100 ml500 ml P04-67100P04-67500   Storage 2° C to ambient temperature „Protect from light!“ When properly stored, separating solutions are stable for at least 36 months. The storage period starts with the manufacturing date. References Nicotinamide phosphoribosyltransferase leukocyte overexpression in Graves’ opthalmopathy (Sawicka-Guta et al.)http://link.springer.com/article/10.1007/s12020-015-0855-8Automated nanoscale flow cytometry for assessing protein–protein interactions (Von Kolontja et al.)http://onlinelibrary.wiley.com/doi/10.1002/cyto.a.22937/fullEstablishment of non-irradiated GVHD mice model (Matsuda et al.) https://www.jstage.jst.go.jp/article/cytometryresearch/26/1/26_D-16-00002/_pdfA Triple Co-Culture Model of the Human Respiratory Tract to Study Immune-Modulatory Effects of Liposomes and Virosomes (Blom et al.)http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0163539Influenza A Virus Infection in Pigs Attracts Multifunctional and Cross-Reactive T Cells to the Lung (Talker et al.)http://jvi.asm.org/content/90/20/9364.shortRegulatory B10 cells display an altered homoeostasis in acute graft-versus-host disease (chakupurakal et al.)http://onlinelibrary.wiley.com/doi/10.1111/ejh.12810/fullMonoclonal antibody 1.6.1 against human MPL receptor allows HSC enrichment of CB and BM CD34+CD38− populations (Cocault et al.) http://www.sciencedirect.com/science/article/pii/S0301472X15008012Rituximab induces phenotypical and functional changes of NK cells in a non-malignant experimental setting (Merkt et al.)https://arthritis-research.biomedcentral.com/articles/10.1186/s13075-016-1101-3Features of Human CD3+CD20+ T Cells (Depke et al.)http://www.jimmunol.org/content/197/4/1111.shortSpecific phenotype and function of CD56-expressing innate immune cell subsets in human thymus (Gerstner et al.)http://www.jleukbio.org/content/early/2016/06/28/jlb.1A0116-038R.shortComparative phenotypical analysis of B cells in fresh and cryopreserved mononuclear cells from blood and tissue of rhesus macaques (Klippert et al.)http://www.sciencedirect.com/science/article/pii/S0022175916300527Effect of chaetocin on renal cell carcinoma cells and cytokine-induced killer cells (Rombo et al.)https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4836455/Der Einfluss physiologischer Sauerstoffkonzentrationen auf natürliche Killerzellen bei der Hepatitis C Virus und der Humanes Immundefizienzvirus Infektion (Franziska Wolter)http://hss.ulb.uni-bonn.de/2016/4406/4406.pdfValidation of an IFNγ/IL2 FluoroSpot assay for clinical trial monitoring (Körber et al.)https://translational-medicine.biomedcentral.com/articles/10.1186/s12967-016-0932-7Yeast (Saccharomyces cerevisiae) Polarizes Both M-CSF- and GM-CSF-Differentiated Macrophages Toward an M1-Like Phenotype (Seif et al.)http://link.springer.com/article/10.1007/s10753-016-0404-5   Separating Solutions Pre-Filled Pancoll separating solutions from PAN-Biotech are made from a neutral, highly cross-linked, hydrophilic polymer of sucrose with an average molecular weight of 400,000 daltons. Pancoll is suited for separation of lymphocytes and other cell types. The ready-to-use solutions are available in 500 ml bottles (see page 105) as well as in pre-filled ready-to-use tubes with a separating membrane. Stability Pancoll is stable for 36 months at 2 °C to 20 °C. Protect from light! Method of seperation For lymphocyte separation blood is used which has been defibrinated or treated with anticoagulants (Heparin, EDTA, Citrate), and which is diluted with the same volume of a physiological saline solution. Then the Pancoll solution is carefully covered with a layer of diluted blood in a centrifuge vial, without mixing the phases. After a short centrifugation step (e.g. 800-1000x g for 20-30 minutes without brake) at room temperature the lymphocytes, together with monocytes and platelets, can be harvested from the white blood cells layer between the plasma sample layer and the Pancoll. The separated cells are then washed twice in physiological saline solution to purify the lymphocytes by removing platelets. During centrifugation the cells of the blood sample migrate to the Pancoll layer where they get into contact with the polysaccharide contained in Pancoll. The red blood cells are aggregated by this substance at room temperature immediately. Aggregation causes an increase of the sedimentation rate of the red blood cells which aggregate together with the granulocytes as a sediment at the bottom of the centrifuge vial. Lymphocytes, monocytes and platelets are not so dense and can not enter and pass through the Pancoll layer. These cells are concentrated as white blood cell layer above the Pancoll layer and therefore can be harvested easily by careful pipetting. In subsequent centrifugation steps the lymphocytes are washed to remove remaining platelets, serum and Pancoll. As a result of this process a highly purified suspension of viable lymphocytes and monocytes (PBMC) is obtained. Description Size Product number Pancoll human, density 1.077 g/ml 25 x 50 ml50 x 10 ml P04-60125P04-60225 Pancoll animal, density 1.077 g/ml 50 x 10 ml P04-63225