Human IGF-I/IGF-1 Quantikine ELISA Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.	-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Upstream tips
-Store the unopened kit at 2-8 °C. -Bring all reagents to room temperature before use. -Reconstitute the Human Cytochrome c Standard with deionized or distilled water. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions.
Protocol tips
-When mixing or reconstituting protein solutions, always avoid foaming. -Substrate Solution should remain colorless until added to the plate. -Keep Substrate solution protected from light. -Stop Solution should be added to the plate in the same order as the Substrate Solution. -Wells that are green in color indicate that the Stop Solution has not mixed thoroughly with the Substrate Solution.

Publication protocol

Stool samples were analysed for calprotectin (Bühlmann fCAL ELISA; Schönenbuch, Switzerland) as a biomarker of intestinal inflammation. Faecal α1-antitrypsin (ELISA, Immunodiagnostik AG; Hamburg, Germany), plasma intestinal fatty acid binding protein (IFABP; Human FABP2/I-FABP Quantikine ELISA Kit; R&D Systems, Inc.; Minneapolis, USA) and plasma IgG anti-endotoxin antibody concentration (EndoCab Human, ELISA; Hycult biotech; Uden, The Netherlands) were measured as markers of intestinal mucosal integrity. Plasma C-reactive protein and α1-acid glycoprotein (Quantikine ELISA, R&D Systems, Inc.; Minneapolis, USA) were measured as markers of acute and chronic systemic inflammation respectively31. Plasma insulin like growth factor (IGF)-1 and IGF binding protein 3 (IGFBP3; Quantikine ELISA, R&D Systems, Inc.; Minneapolis, USA) as growth factors that are depressed in inflammation and SAM were also measured14,32. All analyses followed the manufacturer’s instructions. Normal ranges were those recommended by the manufacturers based on studies in healthy people. Laboratory staff was blinded to treatment allocations.

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Manufacturer protocol

Download the product protocol from R&D Systems for Human IGF-I/IGF-1 Quantikine ELISA Kit below.

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