Immunomagnetic cell isolation is a cost-effective and gentle method of obtaining purified cell populations. However, isolating novel or rare cell populations often relies on relatively time consuming and expensive cell sorting techniques due to the lack of off-the-shelf immunomagnetic cell isolation kits. To bridge this gap, we have developed the EasySep™ Release isolation kits for PE- or biotin-conjugated antibodies. EasySep™ Release uses magnetic particles with low non-specific binding characteristics that can be rapidly removed from isolated cells after positive selection. The system is highly customizable, targeting cells through the use of any biotin or PE-conjugated primary antibody or ligand. Starting with a variety of sample materials, including single cell suspensions from mouse spleen, lymph nodes, lungs, rat spleen, or human PBMCs, the EasySep™ Release kits have demonstrated purities consistently above 90%. The EasySep™ Release technology can also be used to isolate subsets of cells when used in combination with our other cell separation products. For example, functional and particle-free mouse CD25+ or CD304+ CD4+ regulatory T cells were isolated by using our EasySep™ PE Positive Releasable Rapidspheres™ kit and our EasySep™ Mouse CD4+ T Cell Isolation kit. We have used EasySep™ Release with EasySep™ Dextran Rapidspheres™ to isolate CD4/CD8 double positive mouse thymocytes (double positive selection), and to isolate mouse lung epithelial cells by EpCAM positive selection followed by the removal of contaminating leukocytes via CD45 depletion. These kits offer a customizable, and cost-effective magnetic isolation approach for the isolation of virtually any cell population.
Over the course of a lifetime, memory B cells are key to maintaining antibody-mediated protection from viral and bacterial pathogens and they are one outcome of an effective vaccine. Since memory B cells are typically found at frequencies of less than 5% of normal human peripheral blood mononuclear cells (PBMC), research into their function would be aided by a fast and easy isolation method. To meet this need, we have developed an improved EasySep™ kit for the isolation of CD27-positive (CD27+) human memory B cells from fresh PBMC. First, CD27+ cells are labeled using an antibody cocktail and EasySep™ Releasable RapidSpheres™ and are positively selected using a hand-held EasySep™ magnet. Next, magnetic particles are released from the positively-selected cells, and then non-B cells within the CD27+ fraction are targeted for depletion using a second antibody cocktail and EasySep™ Dextran RapidSpheres™. Following an additional magnetic separation step, the particle-free, CD27+ memory B cells are poured off into a new tube and ready for use. With one additional step, CD27− naive B cells can be isolated from the same starting sample. The entire isolation protocol is completed in under 40 minutes without any centrifugation steps. Using this method, CD19+CD27+ memory B cells can be isolated to 97±2% purity and 37±14% recovery (average of n=19 ± S.D.). The CD19+CD27− naive B cells can be isolated to 93±5% purity and 28±11% recovery (n=9 ± S.D.). When stimulated with CpG and IL-15, the isolated memory and naive B cells are functional, as assessed by both proliferative responses and the secretion of IgG.
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