Xvns 15

Manufactured by Lonza

The XVNS-15 is a laboratory equipment designed for performing vacuum-based processes. It features a 15-liter chamber capacity and is suitable for a range of applications that require controlled vacuum conditions.

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Lab products found in correlation

Phytohemagglutinin (pha), by BioLegend (1 mentions) Phytohemagglutinin (pha), by Merck Group (1 mentions) Anti cd4 pe cy7, by BD (1 mentions) Bv421, by BD (1 mentions) Cd1c pe, by Miltenyi Biotec (1 mentions) Facs influx sorter, by BD (1 mentions)

2 protocols using xvns 15

PBMC Cytokine Profiling in Response to LPS and PHA

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PBMC were cultured in 96-well plates (10⁵ cells/well) in serum-free medium (XVNS-15; Lonza), with or without ultrapure LPS from Salmonella enterica serovar minnesota mutant R595 (10 ng/ml; Invitrogen) or PHA (10 μg/ml; Sigma-Aldrich). Cell-free tissue supernatants were collected at 18 h and stored at −20 °C for batched analysis. Cytokine quantifications in cell-free culture supernatants at 1 : 2 dilution were performed using a customized multiplex cytokine array, including: IFN-α2, IFN-γ, IL-10, IL-12p70, IL-13, IL-15, IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, MCP-1, and TNF-α (Luminex 200 System, EMD-Millipore-Sigma). IL-8 levels were above the highest standard in the multiplex array, and retested using aliquoted culture supernatant by commercial ELISA (BioLegend) at a 1 : 20 (rest) or 1 : 40 (LPS/PHA) dilution according to manufacturer's instructions.

Underwood M.L., Nguyen T., Uebelhoer L.S., Kunkel L.E., Korthuis P.T, & Lancioni C.L. (2019). Altered monocyte phenotype and dysregulated innate cytokine responses among people living with HIV and opioid-use disorder. AIDS (London, England), 34(2), 177-188.

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Enrichment and Isolation of Naive CD4+ T Cells

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Cryopreserved CBMC and PBMC were thawed in the presence of DNAse, assessed for viability using trypan blue exclusion, and rested overnight in serum-free complete medium (XVNS-15; Lonza) plus low dose IL-2 (30 U/ml; Prometheus) in 6 well ULA tissue culture plates (Fisher Scientific). Following overnight rest, CBMC/PBMC were subjected to CD4+ enrichment using negative isolation columns (Miltenyi), and resulting CD4+ T cells labeled with anti-CD4-PECy7 (BD), anti-CD45RA-allophycocyanin (Biolegend), and anti-CD14/CD19/CD123/CD56 (Biolegend)/CD1c -PE (Miltenyi), and sorted to obtain CD4+CD45RA+(CD14/CD19/CD123/CD1c/CD56)^negative T cells using a FACS InFlux sorter (BD). To assess the impact of contaminating CD4+CD45RA+RO+ and CD4+CD45RO+RA− T cells on the effector cytokine response among adult donors, PBMC from a subset of adult donors underwent CD4+ enrichment using negative isolation columns followed by parallel FACS purification for CD4+/CD45RA+/CD14, CD19, CD123, CD1c, CD56^negative T cells and CD4+/CD45RA+/CD45RO− (BV 421; BD)/CD14, CD19, CD123, CD1c, CD56^negative T cells (Supplemental Figure 1 A/B).

Sinnott B.D., Park B., Boer M.C., Lewinsohn D.A, & Lancioni C.L. (2016). Direct TLR-2 Co-stimulation Unmasks the Pro-inflammatory Potential of Neonatal CD4+ T cells. Journal of immunology (Baltimore, Md. : 1950), 197(1), 68-77.

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