Ovalbumin Immunization with Adjuvants

Ovalbumin (Catalog number A5503, Sigma-Aldrich, St. Louis, MO) was decontaminated of lipopolysaccharide using a Triton X-114 detoxification method^{60 (link)} and tested with Pierce LAL chromogenic endotoxin quantitation kit (catalog number 88282, Thermo Fisher Scientific, Waltham, MA). Ovalbumin was labeled with Alexafluor 488 using Alexafluor 488 succimidyl ester labeling system (Catalog number A20100, Thermo Fisher Scientific, Waltham, MA). Ovalbumin was used in combination with poly(I:C) (catalog number VAC-PIC, Invivogen, San Diego, CA), and anti-CD40 (clone FGK4.5, BioXCell, West Lebanon, NH)^{61 (link),62 (link)}. For pathogen challenge, female C57BL/6 mice were injected with either 1 × 10⁷ pfu/mouse of vaccinia virus western reserve strain expressing the Ovalbumin protein (VV-ova) intraperitoneally or 1 × 10⁴ pfu/footpad^{24 (link)} using the vaccinia virus western reserve strain with Ovalbumin labeled with Alexafluor 488. For anti-PDPN injection purified clone 8.1.1⁴ (catalog number 127402, Biolegend, San Diego CA) was injected at 100 μg intravenously 4 h before ova-488 and 72 h later.

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Kedl R.M., Lindsay R.S., Finlon J.M., Lucas E.D., Friedman R.S, & Tamburini B.A. (2017). Migratory dendritic cells acquire and present lymphatic endothelial cell-archived antigens during lymph node contraction. Nature Communications, 8, 2034.

Publication 2017

Ovalbumin Pierce LAL chromogenic endotoxin quantitation kit anti-CD40 (clone FGK4.5,

C57bl mice Chromogenic Clone Detoxification Endotoxin Ester Female Lipopolysaccharide Mouse Ovalbumin Pathogen Poly i c Protein Strain Triton x 114 Vaccinia virus

Corresponding Organization : University of Colorado Anschutz Medical Campus

Other organizations : National Jewish Health, University of Colorado Denver

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Protocol cited in 3 other protocols

Variable analysis

independent variables

Ovalbumin (Catalog number A5503, Sigma-Aldrich, St. Louis, MO)
Poly(I:C) (catalog number VAC-PIC, Invivogen, San Diego, CA)
Anti-CD40 (clone FGK4.5, BioXCell, West Lebanon, NH)
Vaccinia virus western reserve strain expressing the ovalbumin protein (VV-ova)
Vaccinia virus western reserve strain with ovalbumin labeled with Alexafluor 488
Anti-PDPN (clone 8.1.1, catalog number 127402, Biolegend, San Diego CA)

dependent variables

Not explicitly mentioned

control variables

Female C57BL/6 mice

positive controls

None specified

negative controls

None specified

Annotations

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