Prostaglandin e2 elisa kit

The level of PGE2 production was used to serve as determining COX activity. Determination of PGE2 levels in serum was performed by Prostaglandin E2 ELISA kit following the manufacturer's protocol (Abcam; cat. no. ab133021). A mouse IgG antibody was precoated onto 96-well plates. Serum samples were added to the wells, along with an alkaline phosphatase (AP) conjugated-PGE2 antibody. After reagent incubation, substrate was added and catalyzed by AP to produce a yellow color. The intensity of the yellow coloration was inversely proportional to the amount of PGE2.

RAW264.7 cells were added to 12-well plates (2 × 10⁶ per well), and
non-adherent cells were removed 2 hr later. Adherent cells were cultured in medium
containing HT (1.56−12.5 µg/ml) with LPS (0.25
µg/ml) at 37°C for 3 or 24 hr. The cells cultured in
the absence of LPS and HT served as controls. The cultured cells were harvested, and
real-time reverse transcription polymerase chain reaction (RRT-PCR) was used to measure
the levels of mRNA-encoding proteins involved in inflammation. The NO, PGE₂,
TNF-α, and IL-1β concentrations in cell culture media harvested following 24 hr of
incubation were determined using a Nitric Oxide (total) detection kit (Enzo Life Sciences,
New York, NY, U.S.A.), Prostaglandin E2 ELISA kit (Abcam, Cambridge, U.K.), Mouse TNF
alpha SimpleStep ELISA^® kit (Abcam), and IL-1 beta Mouse SimpleStep
ELISA^® Kit (Abcam), respectively. The cellular COX-2 concentrations were
determined using a Mouse COX2 SimpleStep ELISA^® kit (Abcam). For western
blotting, the cells were harvested after 15 min to analyze the expression of
phosphorylated NF-κB (pNF-κB) and ERK1/2 (pERK1/2), and after 6 hr to analyze the
expression of iNOS.