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TNF-α, IL-10, and IL-6 were measured using human-specific Luminex™ beadsets (BioSource-Invitrogen, Austin, TX) using a Luminex™ 100 IS apparatus (MiraiBio, Austin, TX) or commercially available, pig-specific ELISA kits (R&D Systems, Minneapolis, MN) using a standard ELISA reader.
The absorbance (450 nm) of each sample was analyzed using a standard ELISA plate reader.
Finally, plates were read at 570 and 620 nm wavelengths using a standard ELISA microplate reader.
Levels of NKp30-Ig or NKp46D2-Ig fusion proteins in the serum were analyzed using a standard ELISA assay.
Absorbance was recorded at 492 nm using a standard ELISA 96-well plate reader (Bio-Rad MPM III 1.133).
Viability was assessed 120 h later by Celltiter96 Aqueous1 (Promega Italia, Milan, Italy) using a standard ELISA reader.
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Serum was eluted from the DBS as described [ 33] and tested using a standard CMV ELISA assay (Sera Quest®, Doral, Florida).
This was at least an order of magnitude better than that observed using a standard microtiter plate ELISA with the same Ab reagents.
Cultures were fed with fresh media on days 4 and 7 and HIV CA-p24 was determined in culture supernatant on day 10 using a standard CA-p24 ELISA assay.
Plasma NfH levels were measured in duplicates with the analyst being blinded to all other information using a standard in-house ELISA [ 27].
TNF-α, interleukin 1β (IL-1β), and IL-6 levels in cell supernatants were measured using a standard quantitative sandwich ELISA (eBioscience, Vienna, Austria).
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