Question: Is there a recommended protocol for isolating PBMCs?
Answer: Unfortunately, we do not have an in house protocol for isolating PBMCs. We purchase fresh human PBMCs from AllCells (https://www.allcells.com/products/peripheral-blood-mononuclear-cells-pbmc). AllCells collect mononuclear cells with a clinical-grade apheresis system (Spectra Optia® Apheresis System) and subsequently deplete RBCs and platelets with ammonium chloride and centrifugation, respectively. Customers have also reported success using the Heparin tubes for blood sampling. We have not tested this in house, but if using this solution, cells should be washed to remove any residual contaminants with PBS + 0.04% BSA. Also, using the highest concentration of cells that go into the sample master mix minimizes carryover of residual heparin.
Products: Single Cell 3', VDJ