Mouse lcn2 duoset elisa kit

The IL-6 and NGAL concentrations in the relevant supernatant or serum were quantified with the Mouse IL-6 DuoSet ELISA Kit and Mouse Lcn2 DuoSet ELISA Kit following the manufacturer’s instructions (R&D Systems).

Levels of fecal lipocalin-2 are considered a more sensitive marker of low-grade intestinal inflammation compared to histological analysis (39 (link)). Lipocalin-2 levels were measured in 100-mg/ml feces (reconstituted in PBS 0.1% Tween 20) using Mouse Lcn-2 DuoSet ELISA kit (R&D Systems, Abingdon, UK) as described previously (40 (link)).

At the end of treatment periods of acute and chronic UC experiments, fecal samples were collected by taking individual mice out of their cages and collecting 7 fecal pellets per mouse in sterile cryogenic vials, which were snap-frozen on dry ice immediately, and stored at −80°C until further processing. Quantification of fecal Lcn-2 was performed as described by Chassaing et al. (2012) [28 (link)] with slight modifications. Briefly, 100 mg of frozen fecal samples were reconstituted in 1 ml of PBS (pH. 7.4) supplemented with 0.1% (v/v) Tween 20, which were homogenized for 3 min using a 1600 MiniG® homogenizer (SPEX SamplePrep, Inc., Metuchen, NJ) to obtain a homogenous fecal suspension. The suspension was centrifuged at 12,000 rpm at 4°C for 10 min and the clear supernatant was used to quantify Lcn-2 levels using the mouse Lcn-2 DuoSet ELISA kit (R&D Systems, Minneapolis, MN) following manufacturer’s instructions. The level of fecal Lcn-2 was expressed as Lcn-2 (ng)/feces (g).