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Anti mouse cd19 apc clone 1d3

Manufactured by BD

Anti-mouse CD19-APC (clone 1D3) is a laboratory reagent used for the detection and analysis of mouse CD19-positive cells. It is conjugated with the fluorescent dye Allophycocyanin (APC) for flow cytometric applications.

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2 protocols using anti mouse cd19 apc clone 1d3

1

Comprehensive Immune Cell Profiling of Murine Lungs

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Lung lobes were digested with 20 μg/ml of collagenase D at 37 °C for 30 min, and the reaction was stopped by the addition of 5 mM of EDTA. Digested lung tissue was passed through 40 μm nylon membrane filter. Spleen and lymph nodes were passed through a 40 μm nylon membrane filter directly. Red blood cells were lysed by treating with ACK lysing buffer (Quality Biological 118-156-101). Single cell suspensions were surface stained at a 1:50 dilution in FACS Buffer (PBS, 10% FBS, 0.1% NaN3 sodium azide) with anti-mouse CD3-APC (clone 17A2; BD Pharmingen), anti-mouse CD19-APC (clone 1D3, BD Pharmingen), anti-mouse CD11b-APC-Cy7 (clone M1/70; BD Pharmingen), anti-mouse CD11c-BUV395 (clone HL3; BD Horizon), anti-mouse Ly6G-AF700 (clone 1A8; BD Pharmingen), anti-mouse Ly6C-PECy7 (clone AL-21; BD Pharmingen), anti-mouse SiglecF-PE-CF594 (clone E50-2440; BD Horizon), anti-mouse MHC-II (clone M5/114.15.2; BD Horizon), anti-mouse CCR2 (clone SA203G11; Biolegend) and anti-mouse CD301a (clone LOM-8.7; Biolegend). Samples were acquired using the LSRFortessa X-20 and analyzed using FlowJo software.
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2

Flow Cytometric Analysis of NS3-Specific CD8+ T Cells

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The frequency of NS3-specific CD8+ T cells was analysed by ex vivo staining of splenocytes using the recombinant soluble dimeric mouse H-2D(b):Ig fusion protein (BD Biosciences, San Jose, California, USA) as described previously.21 29 (link) In brief, 1×106 spleen cells were resuspended in PBS/1% FBS (FACS buffer) and incubated with Fc-blocking antibodies. Cells were then washed and incubated for 90 min with H-2D(b):Ig preloaded with a NS3-derived major histocompatibility complex (MHC) I peptide (eg, NS3 cytotoxic T lymphocyte (CTL) epitope with the amino acid sequence APPPSWDAM, H-2Db). Thereafter, cells were washed and incubated for 30 min with a PE-conjugated rat antimouse IgG1 antibody. Cells were then washed and incubated for 30 min with APC-conjugated rat antimouse CD19 and FITC-conjugated rat antimouse CD8 antibodies. A total of 150 000 events from each sample were acquired on a FACSVerse flow cytometer (BD Biosciences) and analysed using the FlowJo V.9.2 software (Ashland, Oregon, USA). The following antibodies were used: antimouse CD16/32 ‘Fc block’ and antimouse CD19-APC ‘clone 1D3’ (BD Biosciences), and antimouse CD8-FITC ‘clone KT15’ (ProImmune).
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