This chapter describes the preparation of PBMCs from whole blood using a density gradient/cell culture protocol. This includes collection of the PBMC culture media and preparation of cell extracts in order to provide the raw materials for proteomic analyses....
(B) Cell purity was measured before (Start) and after isolation (Easy 250, Purple) based on viable cells. Cells were counted and analyzed by flow cytometry. Data shown as mean ± SD; n = 7 - 9. Figure 8. Typical EasySep™ Direct Protocol View More Protocols and Documentation Find ...
We recommend that you optimize the protocol for your cell type. Day -4: Seed PBMCs 4 days before transduction, thaw PBMCs and gently transfer the cells into one well of a 24-well culture plate. Add 1 mL of StemPro-34 Medium to the cells whil...
2006). This methodology is very efficient in removing target cells due to its fast cell capture kinetics, whereas the use of secondary-antibody coated beads makes it a flexible protocol to isolate any cell of interest (Neurauter et al. 2007). 16.4.2 Indirect Positive Isolation of Human CD4+...
16.4General Protocol 16.4.1T Cell Isolation Protocols Because of the ease of access to peripheral blood, PBMCs, which contain T and B lymphocytes and monocytes, are the major source of human T cells used in most studies. There is a variety of available techniques for isolation and enrichment...
(B) Cell purity was measured before (Start) and after isolation (Easy 250, Purple) based on viable cells. Cells were counted and analyzed by flow cytometry. Data shown as mean ± SD; n = 7 - 9. Figure 8. Typical EasySep™ Direct Protocol View More Protocols and Documentation Find ...
(155 mM NH4Cl; 10 mM KHCO3 and 0.1 mM EDTA) for 5.7 minutes at room temperature. Centrifuge the granulocytes at 300~g for 10 minutes at 20 ‹C. Wash twice with buffer and centrifuge at 200~g. Resuspend the cell pellet in buffer supplemented with BSA or cell culture medium...
Antigen-Specific T-Cell Activation & Expansion Here's a general protocol: Initial Activation: Co-culture PBMCs with APCs loaded with the specific antigen. Expansion: After initial activation, expand T-cells in the presence of IL-2 or other growth factors. ...
Avoid antibody capping on cell surfaces and non-specific cell tagging by working fast, keeping cells and solutions cold through the use of pre-cooled solutions, and by adhering to the incubation times and temperatures specified in the protocol. Increased temperature and prolonged...
We recommend that you optimize the protocol for your cell type. Day -4: Seed PBMCs 4 days before transduction, thaw PBMCs and gently transfer the cells into one well of a 24-well culture plate. Add 1 mL of StemPro-34 Medium to the cells whi...