Anti cd138 biotin

Cells were treated with 5 mM EDTA in PBS before staining, which reduced the binding of cytophilic nonendogenous Abs to B cells (Supplemental information). Cells were treated with unlabeled anti-CD16/CD32 (2.4G2, eBioscience) in FACS buffer (1% BSA/PBS) to block FcγRs. After washing, cells were stained with the relevant antibodies: anti-Igκ-FITC (187.1, BD Pharmingen), anti-IgM-PerCP-Cy5.5 (II/41, eBioscience), anti-IgD-FITC (11–26 c.2a, BD Pharmingen), anti-IgG1-PE (A85-1, eBioscience), anti-CD45R/B220-PE-Cy7 (RA3-6B2, eBioscience), anti-CD138-Biotin (281.2, BD Pharmingen), anti-GL7-FITC (GL7, BD Pharmingen), anti-CD38-PE (90, eBioscience), anti-CD95/Fas-PE (Jo2, BD Pharmingen), anti-CXCR4-APC (2B11, eBioscience), and PE–streptavidin (eBioscience). Cells were analyzed on FACSCanto II and FACSAria II (BD Biosciences). Cell sorting was performed on FACSAria II (BD Biosciences). The data were analyzed using FlowJo software (Tree Star).

Suspensions of single lymphocytes from spleens and Payer’s patches were prepared and stained with antibodies as follows: anti-CD3-pacific blue, anti-CD19-PE/Cy7, anti-CD38-APC, anti-CD138-biotin, anti-Fas-PE/CF594, anti-GL7-FITC and Streptavidin-PE from BD Biosciences (California, USA). The lymphocytes from bone marrows were isolated and stained with anti-CD3-pacific blue, anti-CD19-PE/Cy7, and anti-CD138-biotin. Flow cytometry data were acquired by the flow cytometer (CantoII; BD Biosciences) and analyzed with FlowJo software (version 10; Tree Star Inc, Oregon, USA).