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As a global market leader with numerous subsidiaries and distributors, Miltenyi Biotec is committed to providing our customers around the world with the highest quality products. In addition to direct selling in more than 20 countries in North America, Europe and Asia/Pacific, Miltenyi Biotec also provides support for our customers through an extensive distributor network covering dozens of additional countries.
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This application protocol describes an exemplary procedure, which could be used in order to isolate LOX-1+ cells. Here, LOX-1 cells were isolated from cord blood mononuclear cell suspension from one individual.
The human LOX-1 antigen, the lectin-like oxidized low-density lipoprotein receptor 1, is also known as OLR1 or CLEC8A and belongs to the C-type lectin superfamily. LOX-1 is found on endothelial cells, macrophages, platelets, fibroblasts, and smooth muscle cells. It plays a role in endocytosis, phagocytosis, apoptosis, and inflammatory processes. LOX-1 is expressed on a population of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) in peripheral blood of cancer patients1. Moreover, a high density of tumor-associated neutrophils (TANs) expressing LOX-1 and arginase 1 is strongly correlated with a reduced expression of cytotoxic granzyme B and the proliferation marker Ki67 in closely located T cells, suggesting a suppressive function of LOX-1+ TANs2.
When working with anticoagulated cord blood, cord blood mononuclear cells should be isolated by density gradient centrifugation, e.g., using Ficoll-Paque™. For details regarding the preparation of mononuclear cells please refer to the Resources section of the Diamond CD34 Isolation Kit, human.
▲Note: To remove platelets after density gradient separation, resuspend cell pellet in buffer and centrifuge at 200×g for 10−15 minutes at 20 °C. Carefully aspirate supernatant. Repeat washing step.
▲Note: It is strongly recommended to screen biological samples for the presence of LOX-1+ cells before proceeding with the separation protocol.
1.1 Labeling of LOX-1+ cells using LOX-1 Antibody, anti-human, PE, REAfinity™
1.2 Labeling of LOX-1+ cells using LOX-1 Antibody, anti-human, APC, REAfinity
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