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INTENDED USE The BD Vacutainer® CPT™ Cell Preparation Tube with Sodium Citrate is an evacuated tube intended for the collection of whole blood and the separation of mononuclear cells. The cell separation medium is comprised of a polyester gel and a density gradient liquid. This configuration permits cell separation during a single centrifugation step. The separated sample can be transported without being removed from the BD Vacutainer® CPT™ Tube since the gel forms a stable barrier between the cell layers. SUMMARY AND EXPLANATION Isolation of mononuclear cells from whole blood is a first step for many in vitro assays. One currently accepted technique for mononuclear cell separation, referred to as the FICOLL™ Hypaque™ method, employs a liquid density gradient medium of FICOLL™ 400 and sodium metrizoate or sodium diatrizoate solution.1,2,3 The procedure uses anticoagulated blood, collected by routine phlebotomy, which is diluted with a buffered solution, and then carefully layered onto the medium. This preparation is then centrifuged to isolate the mononuclear cells above the medium. The cells are harvested by carefully pipetting them from the liquid interface. The BD Vacutainer® CPT™ Cell Preparation Tube with Sodium Citrate combines a blood collection tube containing a citrate anticoagulant with a FICOLL™ Hypaque™ density fluid and a polyester gel barrier which separates the two liquids. The result is a convenient, single tube system for the collection of whole blood and the separation of mononuclear cells. The BD Vacutainer® CPT™ Cell Preparation Tube with Sodium Citrate reduces the risk of sample contamination and eliminates the need for additional tubes, pipettes, and reagents. Samples can be transported without removing them from the tube. TEST PRINCIPLES The BD Vacutainer® CPT™ Cell Preparation Tube with Sodium Citrate is an evacuated blood collection tube system containing 0.1 M sodium citrate anticoagulant and blood separation media composed of a thixotropic polyester gel and a FICOLL™ Hypaque™ solution. The blood separation media takes advantage of the relatively low density of mononuclear cells to isolate them from whole blood. The separation occurs during centrifugation when the gel portion of the medium moves to form a barrier separating the mononuclear cells and plasma from the denser blood components. The mononuclear cells can be collected by pipetting the cell layer, or the cells can be resuspended into the plasma by gentle inversion to improve cell viability if the sample is to be transported
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文献和实验Preparation of Carboxypeptidase Y and Its Properties
to autodigestion. Carboxypeptidase Y is stored as zymogen in yeast. It can be separated from other proteins in yeast by the above-described properties. First, the yeast cell autolyze with organic solvents, and activate the enzyme under pH5.0 in order
Human Peripheral Blood Mononuclear Cell Preparation
in 1 to 2 ml of PBS, and transfer the latter to a fresh tube (to avoid contaminating the solution with the platelets that are adhering to the sides of the tube).7. Bring the cell suspension to a final volume of 50 ml and gently overlay 12.5 ml onto the Ficoll
Native Chromatin Preparation and Illumina/Solexa Library Construction
is empty. Collect the entire effluent in the same tube (from Step 18). 20. Count cells and check CD4+ T cell purity. Recovery is typically ~20%. Purified CD4+ T cells with ~ 95%-98% purity are used for native chromatin preparation
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