Ovalbumin (OVA) was purchased from Sigma. Monosodium urate and basic calcium phosphate crystals, both used at 250 ug/ml for antigen presentation assays, were made from materials from Sigma as described previously [33 (link), 37 (link)]. For MSU, the crude crystal preparation was tumbled with 5 mm stainless steel beads in a 50 ml conical tube for one week to produce finer crystals about 1um in length. Latex beads (3 um) were purchased from Polyscience Inc. Complete Freund’s adjuvant (CFA) and incomplete Freund’s adjuvant (IFA) were from Rockland. All IL-2 ELISA Ready-Set-Go kits were from eBioscience. Alexa-488 OVA, Lysotracker (deep red and green) and Alexa-594 Transferrin receptor were bought from Invitrogen. Anti mouse CD4 was from BD. Anti mouse antibodies for CD8, CD40, CD45.1, CD69, CD80, CD86, CD90.1, MHC I (H2Db), MHC II (Ia/Ie), IL-4 and IFN-γ were from eBioscience Inc. Secondary antibodies and Prolong Gold Antifade were bought from Molecular Probes (Invitrogen).
Protocol detail
Antigen Presentation Assay Protocol
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Ovalbumin (OVA)
Complete Freund’s adjuvant (CFA)
incomplete Freund’s adjuvant
Anti mouse CD4
Prolong Gold Antifade
Alexa 594
Anti antibodies
Antibodies
Antigen presentation
Assays
Calcium phosphate
Cd90
Elisa
Freund adjuvant
Gold
Ifn γ
Lysotracker
Molecular probes
Monosodium urate
Mouse
Ovalbumin
Stainless steel
Transferrin receptor
Corresponding Organization :
Other organizations : University of Calgary, Center for Life Sciences, Tsinghua University, University of Massachusetts Chan Medical School
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Variable analysis
independent variables
- Ovalbumin (OVA)
- Monosodium urate (MSU) crystals
- Basic calcium phosphate crystals
- Latex beads (3 um)
- Complete Freund's adjuvant (CFA)
- Incomplete Freund's adjuvant (IFA)
- Antigen presentation assays
- Concentration of MSU crystals and basic calcium phosphate crystals (both used at 250 ug/ml)
- Alexa-488 OVA
- Lysotracker (deep red and green)
- Alexa-594 Transferrin receptor
- Anti mouse CD4, CD8, CD40, CD45.1, CD69, CD80, CD86, CD90.1, MHC I (H2Db), MHC II (Ia/Ie), IL-4 and IFN-γ antibodies
- Secondary antibodies
- Prolong Gold Antifade
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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