Elisa technique kits

Serum Tumor Necrosis Factor alpha (TNFα), Interferon gamma (IFN γ), Interlukin 6 (IL6), Interlukin 10 (IL10), Interlukin 1beta (IL1β), Transforming growth Factor beta (TGFβ), Interlukin 1 Receptor antagonist (IL1Ra), Interlukin 18 (IL18), Interlukin 8 (IL8) and C-reactive protein (CRP) levels in serum were measured by using enzyme-linked immunosorbent assay (ELISA) technique kits from Abcam Biotech Co., Cambridge, UK and G-Biosciences, Geno Technology Inc., USA. These assays detected only human cytokines and at very low serum concentrations. ELISA was performed as per manufacturer’s protocol.

The concentrations of tumour necrosis factor (TNF)-α, interferon (IFN)-γ, IFN-β, interleukin (IL)-6, IL-1β, IL-27, IL-17, and IL-10 were measured with a commercially available enzyme-linked immunosorbent assay (ELISA) technique kits following the manufacturer’s recommendations (R&D Systems, Minneapolis, MN, USA). The chemokine (C–C motif) ligand 2 (CCL2) concentration was measured with commercially available ELISA technique kits following the manufacturer’s recommendations (Abcam, Cambridge, UK).

Poly(I:C), the TLR3 agonist, was administered as described previously [20 (link),21 (link)]. Briefly, two days after the last day of treatments with HK36, HK39, or HK40, mice received 100 μL of PBS containing 250 μg poly(I:C) (equivalent to 10 mg/kg body weight) through the nasal route. Control mice received 100 μL of PBS. Animals received three doses of poly(I:C) or PBS with 24 h rest period between each administration.
Broncho-alveolar lavages (BAL) samples were obtained as described previously [17 (link),23 (link)]. Albumin content was determined colorimetrically using an albumin diagnostic kit (Wiener Lab, Buenos Aires, Argentina). BAL lactate dehydrogenase (LDH) activity was assessed with the Wiener reagents and procedures (Wiener Lab).
IFN-β (Mouse IFN-beta ELISA Kit), IFN-γ (Mouse IFN-gamma Quantikine ELISA Kit), IL-6 (Mouse IL-6 Quantikine ELISA Kit), IL-10 (Mouse IL-10 Quantikine ELISA Kit), IL-12 (Mouse IL-12 p70 DuoSet ELISA), tumor necrosis factor (TNF)-α (Mouse TNF-α 236 ELISA Kit) and IL-27 (Mouse IL-27 p28/IL-30 Quantikine ELISA Kit) concentrations in BAL samples were measured with commercially ELISA technique kits following the manufacturer’s recommendations (R&D Systems, MN, USA). CCL2 (Mouse MCP1 ELISA Kit (ab208979), and chemokine KC (or CXCL1) were measured with commercially available ELISA technique kits following the manufacturer’s recommendations (Abcam).