Cd138 bv605

Manufactured by BioLegend

CD138 BV605 is a fluorochrome-conjugated antibody used for the detection and analysis of CD138-expressing cells in flow cytometry applications. CD138, also known as Syndecan-1, is a transmembrane heparan sulfate proteoglycan that is expressed on the surface of plasma cells and certain epithelial cells. The BV605 fluorochrome provides a specific signal for identification and quantification of CD138-positive cells.

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2 protocols using cd138 bv605

Identification of Germinal Center and Plasma Cells

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Single-cell suspensions were obtained by forcing popliteal LNs through a 70-μm mesh into ice-cold FACS buffer (EDTA 1 mM and 2% serum in PBS). Cells were incubated with 2 μg/ml anti-16/32 (clone 93) for blockage of Fc receptors for 5–10 min. Cell suspensions were washed and incubated with fluorescently labeled antibodies (B220 V500, FAS PE/Cy7, CD38 A700, CD138 BV605, CD80 APC, PD-L2 PC5.5; BioLegend) for 20–40 min. GC cells were gated as live/single, B220⁺ CD38^Lo FAS^Hi. PCs were defined as B220^med CD138⁺. TdTomato⁺ cells were detected by the PE channel for AID.Cre.tdTomato and AID.Cre.ERT2.tdTomato. Cell suspensions were analyzed by Cytoflex (Beckman) flow cytometer.

Stoler-Barak L., Biram A., Davidzohn N., Addadi Y., Golani O, & Shulman Z. (2019). B cell dissemination patterns during the germinal center reaction revealed by whole-organ imaging. The Journal of Experimental Medicine, 216(11), 2515-2530.

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Isolation and Analysis of Splenic Dendritic Cells

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Spleen mononuclear cells were prepared as previously described [69 (link)]. For studies of cDCs, spleens were treated with deoxyribonuclease I (0.5mg/ml; Worthington Biochemical) and collagenase type 4 (1mg/ml; Worthington Biochemical) for 25 minutes at room temperature, in order to ensure maximal recovery of splenic cDCs. Fluorescently conjugated monoclonal antibodies, anti-mouse B220-Alexa Fluor 700 (RA3-6B2), B220-Pacific blue (RA3-6B2), CD19-FiTC (6D5), CD138-BV605 (281–2), IgD-APCCy7 (11-26c.2a), IgM-PECy7 (RMM-1), TCRβ-Alexa Fluor 700 (H57-597), TCRβ-APC/Cy7 (H57-597), CD4-BV605 (RM4-5), IFNγ-BV421 (XMG1.2), ICOS-PE (7E.17G9), Streptavidin-PE/Cy7, CD45.1-FiTC (A20), CD45.2-Alexa Fluor 700 (104), CD11c-APC (N418), MHCII-Pacific blue (M5/114.15.2), CD8α-PE/Cy7 (53–6.7), CD40-PE (1C10), CD80-PE (16-10A1), CD86-PE (GL1), ICOSL-PE (B7-RP1), PDL1-PE (MIH5), PDL2-PE (TY25), CD49d-Biotin (R1-2), CD11a-FiTC (M17/4), Ki-67-PE (16A8) and Zombie Aqua fixable viability dye were purchased from Biolegend (San Diego, CA). Anti-mouse CD95/Fas-BV421 (Jo2), CXCR5-biotin (2G8), and Bcl6-PerCP/Cy5.5 (K112-91) were purchased from BD Biosciences (Franklin Lakes, NJ). Anti-mouse T-bet-APC (eBio4B10), GL-7-APC (GL-7) and PD1-APC/Cy7 (J43) were purchased from eBioscience. Cell surface and intracellular IFNγ, T-bet and Bcl6 staining was performed as previously described [69 (link),70 (link)].

Sebina I., James K.R., Soon M.S., Fogg L.G., Best S.E., de Labastida Rivera F., Montes de Oca M., Amante F.H., Thomas B.S., Beattie L., Souza-Fonseca-Guimaraes F., Smyth M.J., Hertzog P.J., Hill G.R., Hutloff A., Engwerda C.R, & Haque A. (2016). IFNAR1-Signalling Obstructs ICOS-mediated Humoral Immunity during Non-lethal Blood-Stage Plasmodium Infection. PLoS Pathogens, 12(11), e1005999.

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