Mouse rat ccl2 je mcp 1 quantikine elisa kit

To determine MCP-1 secretion from human pericytes and murine C₂C₁₂ cells, respective ELISAs for the detection of human (Human CCL2/MCP-1 Quantikine ELISA kit; R&D Systems, Minneapolis, MN) and murine MCP-1 (Mouse /Rat CCL2/JE/MCP-1 Quantikine ELISA kit; R&D Systems) were performed on cell culture supernatants. The kits were tested for species cross reactivity by the manufacturer, and no cross reactivity was detected. Assay kits employed the quantitative sandwich ELISA technique, and were performed on undiluted samples according to manufacturer's instructions. A multiwell microplate reader was used (FLUOstar Optima; BMG Labtech) to measure absorbance at 450 nm. Assays were performed in duplicate and average values were used for analysis.

Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected using ADVIA 2400 Chemistry System (Siemens, Tarrytown, NY). The inflammatory response was measured by detecting the levels of mouse serum cytokines TNF-α, IL-1β, and IL-6 using ELISA kits: murine TNF-α Standard ABTS ELISA Development Kit (900-T54, PeproTech, Rocky Hill, NJ) and mouse/rat Ccl2/JE/MCP-1 Quantikine ELISA Kit (MJE00, R&D Systems, Minneapolis, MN).

Preparation of lung lysate sample for ELIA and immunoblot were described in S1 Appendix. Interleukin (IL)-6, tumor necrosis factor alpha (TNF-α), monocyte chemoattractant protein-1 (MCP-1), and receptor for advanced glycation end products (RAGE) level were measured in serum by ELISA using the Mouse IL-6 Quantikine ELISA kit, Mouse TNF-α Quantikine ELISA kit, Mouse/Rat CCL2/JE/MCP-1 Quantikine ELISA Kit, and Mouse RAGE Quantikine ELISA Kit (all from R&D Systems, Inc., Minneapolis, MN, USA) following the manufacturer's instructions.
SP-A and SP-D were determined from BAL, lung lysate, the kidneys and serum by ELISA. Mouse Sftpa1 ELISA Kit (Aviva Systems Biology) and Mouse SP-D Quantikine ELISA Kit (R&D systems, Minneapolis, MN) were used following the manufacturer’s instructions to measure SP-A and SP-D respectively.