Human Chitinase 3-like 1 DuoSet ELISA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Bring all reagents to room temperature before use. Allow all components to sit for a minimum of 15 minutes with gentle agitation after initial reconstitution. Working dilutions should be prepared and used immediately.	A standard curve should be generated for each set of samples assayed. It is suggested to start Reagent Diluent optimization for serum and plasma samples by using PBS supplemented with 10-50% animal serum. Do not use buffers with animal serum to reconstitute or dilute the Detection Antibody or Streptavidin-HRP B.

Upstream tips
Bring all reagents to room temperature before use. Allow all components to sit for a minimum of 15 minutes with gentle agitation after initial reconstitution. Working dilutions should be prepared and used immediately.
Protocol tips
A standard curve should be generated for each set of samples assayed. It is suggested to start Reagent Diluent optimization for serum and plasma samples by using PBS supplemented with 10-50% animal serum. Do not use buffers with animal serum to reconstitute or dilute the Detection Antibody or Streptavidin-HRP B.

Publication protocol

Maternal peripheral plasma samples were collected and stored in EDTA at -80°C prior to testing. Plasma samples were assessed (as indicated) for levels of Ang1, Ang2, Angiopoietin-Like 3 (AngptL3), Vascular Endothelial Growth Factor (VEGF-A), Factor D, Monocyte Chemoattractant Protein-1 (MCP-1), Interleukin-1 Beta (IL1B), Soluble fms-like tyrosine kinase 1 (sFlt-1), soluble Tumor Necrosis Factor Receptor 2 (sTNFR2), Placental Growth Factor (PGF), Macrophage Inflammatory Protein-1 Beta (MIP-1β), Leptin, Interleukin-18 Binding Protein (IL-18BP), soluble Intercellular Adhesion Molecule-1 (sICAM-1), soluble Endoglin (sEndoglin), C-reactive protein (CRP), Chitinase-3-Like Protein-1 (CHI3L1), and complement component C5a (C5a) using commercially available enzyme linked immunosorbent assay (ELISA) kits (Duosets, R&D Systems, Minneapolis, MN). All laboratory procedures followed the manufacturers’ protocol (R&D System Duosets) with two notable exceptions; our laboratory used a longer incubation period (overnight at 4°C) and samples were developed using ExtrAvidin (1:1000 dilution, Sigma-Aldrich Co.) and SIGMAFAST p-Nitrophenyl phosphate Tablets (Sigma-Aldrich Co.).

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Manufacturer protocol

Download the product protocol from R&D Systems for Human Chitinase 3-like 1 DuoSet ELISA below.

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