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Table 1 Antibody panels used for FCM analyzes

From: PRRSV-infected monocyte-derived dendritic cells express high levels of SLA-DR and CD80/86 but do not stimulate PRRSV-naïve regulatory T cells to proliferate

Antigen Clone Isotype Fluorochrome Labeling strategy Source of primary Ab
Phenotyping of moDCs
CD1 76-7-4 IgG2a Alexa647 secondary antibodya in housej
CD14 CAM36A IgG1 BV421 secondary antibodyb VMRD
CD80/86 CD152/Fc chimera IgG2a Alexa647 secondary antibodya Sigma-Aldrich
CD163 2A10/11 IgG1 PE directly conjugated AbD Serotec
CD169 3B11/11 IgG1 BV421 secondary antibodyb AbD Serotec
CD172a 74-22-15A IgG2b Alexa488 secondary antibodyc in housej
SLA-DR MSA3 IgG2a Alexa647 secondary antibodya in housek
Assessment of moDC infection rate
CD172a 74-22-15A IgG2b PE secondary antibodyd in housej
PRRSV-N P10/b1 IgG1 Alexa488 directly conjugatede in housel
CD163, CD169, SLA-DR and CD80/86 expression on infected moDCs
CD172a 74-22-15A IgG2b Alexa647 secondary antibodyf in housej
SLA-DR MSA3 IgG2a PE directly conjugatedg in housek
CD80/86 hCTLA4/Fc chimera IgG2a BV421 two step biotin-streptavidinh Sigma-Aldrich
CD163 2A10/11 IgG1 PE directly conjugated AbD Serotec
CD169 3B11/11 IgG1 BV421 two step biotin-streptavidini AbD Serotec
PRRSV-N P10/b1 IgG1 Alexa488 directly conjugatede in housel
  1. aGoat anti-Mouse IgG2a-Alexa647, Life Technologies.
  2. bRat anti-Mouse IgG1-BV421, clone RMG1-1, BioLegend.
  3. cGoat anti-Mouse IgG2b-Alexa488, Life Technologies.
  4. dGoat anti-Mouse IgG2b-PE, Southern Biotech.
  5. eAlexa Fluor® 488 Protein Labeling Kit, Life Technologies.
  6. fGoat anti-Mouse IgG2b-Alexa647, Life Technologies.
  7. gLightning-Link™ RPE Conjugation Kit, Innova Biosciences.
  8. hGoat anti-Mouse IgG2a-biot., Southern Biotech; Streptavidin-BV421, BioLegend.
  9. iGoat anti-Mouse IgG1-biot., Southern Biotech; Streptavidin-BV421, BioLegend.
  10. jPescovitz et al. [27].
  11. kHammerberg and Schurig [28].
  12. lWeiland et al. [29].
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Veterinary Research

ISSN: 1297-9716