How to Use SepMate™ to Isolate PBMCs from Whole Blood in Just 15 Minutes [old]

How to Use SepMate™ to Isolate PBMCs from Whole Blood in Just 15 Minutes [old]


[♪ Music ♪] [SepMate™: Hassle-Free PBMCs in Just 15 Minutes] SepMate™ is a new product from STEMCELLTechnologies [Vicki Stronge, PhD. – Product Manager] that lets you isolate PBMCs from whole blood in just fifteen minutes. [Technical Guide] This video demonstrates how to use SepMate™ to isolate PBMCs from whole blood. Start by smoothly pipetting 15 mL of density gradient medium into the SepMate™ tube through the central hole in the insert. Bubbles may be present in the medium but this will not affect the performance. Next, dilute the blood sample with an equal volume of PBS containing 2% FBS. Keeping the SeptMate™ tube in the upright position, pipette the sample quickly down the side of the tube. SepMate™ keeps the sample separated from the density gradient medium. Next, centrifuge the samples at 1200 x g for 10 minutes at room temperature with the brake on. SepMate™ prevents the layers from mixing as the centrifuge slows down. Red blood cells and granulocytes pellet to the bottom of the tube during centrifugation while PBMCs remain above the insert. Some red blood cells may be present on the surface of the insert, but these cells will not affect the purity of your PBMCs. Harvest the PBMCs by pouring the top layer into a new tube in one smooth motion. Do not leave the tube inverted for more than two seconds. SepMate™ keeps red blood cells and density gradient medium separate from the isolated PBMCs. Finally, wash the enriched cells twice. The PBMCs are now ready for downstream applications. SepMate™ can also be used with RosetteSep™, STEMCELL’s one-step cell enrichment system. Using RosetteSep™ with SepMate™ lets you go from whole blood to purified cells in just 25 minutes. To isolate purified cells subsets, simply add the recommended volume of the appropriate RosetteSep™ cocktail to the sample. Mix the sample well and then incubate for 10 minutes at room temperature. Next, simply proceed with the standard SepMate™ protocol. During density gradient centrifugation, unwanted cells pellet along with erythrocytes leaving your cells of interest untouched and highly purified. For more information on SepMate™, RosetteSep™, or other products please visit www.stemcell.com. [www.stemcell.com] [www.sepmate.com] [STEMCELLTechnologies™ Scientists Helping Scientists™] [www.stemcell.com]