Cd19 biotin

Manufactured by BD

CD19-biotin is a lab equipment product that functions as a cell surface marker. It is used to identify and isolate B lymphocytes (B cells) in various research and diagnostic applications.

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3 protocols using cd19 biotin

Comprehensive Mouse Immune Cell Analysis

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Flow cytometric analysis of total mouse splenocytes or bone marrow cells was performed using the following antibodies: B220-BV605 (RA3-6B2), CD4-AF700 (RMP4-5), CD44-APC (IM7), CD62L-PECy7 (MEK-14), PD1-PE (29F.1A12), IgM-BV605 (RMM-1), IgD-BV711 (11-26c2a), CD93-PE (AA4.1), Streptavidin-PECy5, MHCII-PECy7 (M5/114.15.2), Ly51-biotin (6C3), CD24-APC (M1/69), CD23-biotin (B3H4). GL7-FITC (GL-7), CD95- PeCy7, CXCR5-biotin (2G8), CD43-FITC (S7), CD19-biotin (1D3), CD90.2-biotin (53-2.1), CD11b-AF700 (M1/70), CD11c-FITC (HL3) (BD Biosciences). CD8α (clone 53–6.7) (eBioscience). All cells were stained with fixable viability dye-eFluor780 (Invitrogen) prior to surface staining. Stained cells were analyzed using the BD LSR II flow cytometer (BD Biosciences). Data were acquired using FACSDiva software (BD Biosciences) and analyzed using FlowJo software (Tree Star).

Chodisetti S.B., Fike A.J., Domeier P.P., Schell S.L., Mockus T.E., Choi N.M., Corradetti C., Hou B., Atkins H.M., Caricchio R., Decker T., Lukacher A.E., Olsen N, & Rahman Z.S. (2020). Serine phosphorylation of the STAT1 transactivation domain promotes autoreactive B cell and systemic autoimmunity development. Journal of immunology (Baltimore, Md. : 1950), 204(10), 2641-2650.

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Isolation of Murine Immune Cells

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Spleen and lymph nodes were harvested from 6–12-wk-old mice. Single-cell suspension was obtained by dissociation using sterile frosted slides and passing through a 70-µm cell strainer. Red blood cell lysis was performed as needed. Naive CD4 T cells were isolated according to MojoSort kit protocol (BioLegend). The purity of naive CD4 T cells was constantly monitored and maintained at >95% CD4⁺MHCII⁻CD62L⁺CD44⁻. Splenic DCs were isolated from the spleen of a mouse injected with B16-FLT3L melanoma. Splenocytes were blocked with Fc block (anti-CD32/CD16) and stained with CD11c-biotin (BioLegend), then with anti-Biotin beads (Miltenyi), and isolated using AutoMacs magnetic selection (Miltenyi). The purity of isolated splenic DCs was maintained at >98% CD11c⁺. For all experiments, DC donor mice and naive CD4 T cell donor mice were age and sex matched. B cells were isolated from sex- and age-matched naive mouse spleen by isolating the CD19⁺ population using CD19-biotin (BD) and AutoMacs (Miltenyi). LP lymphocytes (LPLs) were isolated as previously described (Hu et al., 2011 (link)).

Gao Y., Deason K., Jain A., Irizarry-Caro R.A., Dozmorov I., Coughlin L.A., Rauch I., Evers B.M., Koh A.Y., Wakeland E.K, & Pasare C. (2020). Transcriptional profiling identifies caspase-1 as a T cell–intrinsic regulator of Th17 differentiation. The Journal of Experimental Medicine, 217(4), e20190476.

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Multiparametric Flow Cytometry Analysis

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Antibodies used to detect the following mouse proteins were: CD4-PerCP, -647, -FITC (RM4-5), CD8-biotin, -PerCP, -647 (53-6.7), CD11b-biotin (M1/70), CD16/32 purified (2.4G2), CD19-biotin (1D3), CD25-PerCP, -APC, -PE (3C7), Gr1-biotin (RB6-8C5), CD44-BV421, -PE and -APC (IM7); TCRβ-APC, -PE, and -BV421 (H57-597); CD3e-PerCP, -APC and -PE -biotin (2C11), NK1.1-biotin (PK136), pTα purified (2F5), CD3γε-APC (17A2), anti-mouse pZAP70 -647 (Y319), obtained from BD Pharmingen; F4/80biotin (BM8), CD98-PE (RL388), CD8-BV421 (53-6.7), CD71-PE (R71217), all from eBioscience. Annexin V-PE, 7AAD and the APC-labeled anti-BrdU mAb (3D4) were purchased from BD Pharmingen. The APA1/1 monoclonal antibody, which recognizes a conformational epitope of CD3e, and its use as a probe for the conformational change of the TCR have been described previously (Risueno et al., 2005) (Risueno et al., 2005) .
Where necessary, secondary antibodies (anti-rabbit Alexa647 and anti-mouse Alexa647 from ThermoFisher) or fluorescent probes (Streptavidin-PErcp and -APC from BD Pharmingen, Streptavidin-PE from Invitrogen and Streptavidin -BV421 from Biolegend) were used.

Bovolenta E.R., García‐Cuesta E.M., Ponomarenko J., Mellado M., Castro M., Abia D., & Santen H.M.v. (2020). Pre-TCR signaling intensity shapes the TCRβ repertoire.

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