Lactoferrin hlf2 human elisa kit

Degranulation was measured by the release of granule proteins by enzyme-linked immunosorbent assay (ELISA). Briefly, neutrophils (0.2 × 10⁶ per well) were suspended in RPMI 1640 containing 10% fetal bovine serum (FBS) in 96-well plates and stimulated with bacteria at 37°C for 4 h. Separately, neutrophils were also stimulated with latrunculin (1 μM) for 30 min, followed by fMLF (N-formylmethionyl-leucyl-phenylalanine) (1 μM) for 10 min, to allow for maximal degranulation. Elastase or lactoferrin release was measured in cell-free supernatants by ELISA (elastase/ELA2 DuoSet ELISA kit from R&D and lactoferrin [HLF2] human ELISA kit from Abcam) following the manufacturers’ instructions.

CTSS activity was determined with the CTSS activity kit (BioVision) according to the manufacturer’s instructions. Activity values were normalised to total protein concentration as described previously^{5 (link)}. sIgA, LF, and Cys C levels in tears eluted from Schirmer’s strips were quantified using the sIgA ImmuChrom ELISA Kit (Eagle Biosciences, Amhurst NH), lactoferrin (HLF2) human ELISA Kit (Abcam), and Cys C quantikine human Cys C ELISA kit (R&D Systems), respectively. Final tear dilutions were 1:5,000 for sIgA, 1:60,000 for LF, and 1:50 for Cys C.