Standard 4 hour 51Cr release assays were performed and percent specific lysis calculated as previously described.6 (link) Unmodified K562 cells were included as a positive control target and the myeloma cell line OPM2 was included when primary myeloma cells were unavailable (American Type Culture Collection, Manassas, VA). Effector cells were freshly cultured (not cryopreserved) expanded NK cells or NK cells enriched from PB samples or BM using microbeads coated with anti-CD56 antibody (Miltenyi Biotec). Research grade expanded NK cells were produced as previously described.6 (link) CD138-positive plasma cells were purified from BM aspirates via positive selection with anti-CD138 coated microbeads (Miltenyi Biotec). After selection, purity of >90% was confirmed by flow cytometry. For blocking studies, isotype control antibody or anti-KIR antibody was added to effector cells. Control antibody, elotuzumab, or anti-HLA Class I antibody was added to 51Cr-labeled targets. Antibodies were left in the wells for the duration of the assay, at a final concentration of 10μg/mL.
Anti cd138 coated microbeads
Manufactured by Miltenyi Biotec
Sourced in United States
Anti-CD138 coated microbeads are a lab equipment product used for the isolation and enrichment of CD138-positive cells from various sample types. These microbeads are coated with antibodies specific to the CD138 cell surface antigen, enabling the magnetic separation and purification of CD138-expressing cells.
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Lab products found in correlation
2 protocols using anti cd138 coated microbeads
1
NK Cell-Mediated Cytotoxicity Assay
2
Isolation and Characterization of Multiple Myeloma Plasma Cells
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Multiple myeloma PCs (MM-PCs) were purified from donated bone marrow aspirates from patients diagnosed with MM (excluding MGUS or SMM)58 (link) using monoclonal antibody anti-CD138-coated microbeads (Miltenyi-Biotec, Auburn, CA, USA). Normal PCs were obtained from discarded bone marrow specimens with no abnormalities at diagnosis after FAC sorting CD138+ CD19+/− CD38+++ cells. Naïve human B-lymphocytes were isolated from peripheral blood samples from normal volunteers after T-cell depletion by a rosette technique and FAC-sorted to obtain CD19+ CD27− IgD+ cells. All human samples (MM n = 7, normal PCs, n = 4, and B-cells, n = 3) were obtained from adult donors upon written informed consent under the supervision and approval of the Institutional Review Board (code PI11/01091, date 11/10/2011, Ethics Committee for Research of the Hospital Universitario “Puerta del Mar”, Cadiz, Spain). The Andalusian Public Health System Biobank coordinated the collection, processing, management and assignment of all biological samples used in this study according to standard procedures established for this purpose.
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