Prostaglandin E2 Assay Kit

Prostaglandin E2 (PGE2) is a primary product of arachidonic metabolism and is synthesized via the cyclooxygenase (COX) and prostaglandin synthase pathways. PGE2 production is a commonly used method for the detection of COX-1 and COX-2 modulation and prostaglandin synthases. The HTRF® PGE2 assay is a highly sensitive method for quantifying PGE2 either in cell supernatants or directly in the presence of whole cells. Our reagents have simplified protocols requiring only two reagent additions. These protocols are easily miniaturized and amenable to automation and HTS.

Assay principle

The PGE2 assay is a competitive immunoassay that uses Eu3+-Cryptate labeled anti-PGE2. Free PGE2 from the sample competes with d2 labeled PGE2 for binding to the Cryptate conjugated anti-PGE2 antibody. The assay can be run in two easy steps, and requires only a single incubation following cell stimulation. Assays can also be run with purified enzymes.

Product specifications

Detection limit: 10 pg/mL
Dynamic range: 8 to 1000 pg/mL
EC50: 250 pg/mL
S/B: 31
Z': 0.9 for 20 µl assay volumes

The PGE2 assay has a high level of  flexibility. It can be performed using cell supernatants or directly on stimulated cells (see figure 1). The incubation time and temperature following addition of the HTRF detection reagents has little effect on the assay results, providing further assay flexibility.

Product performances

One benefit of the HTRF PGE2 assay is its ability to quantify PGE2 in the presence of cells without sacrificing assay performance. The results comparing direct cell-based versus supernatant transfer did not differ significantly. Monocytes were stimulated to produce PGE2 in the presence or absence of indomethacin, a known inhibitor of PGE2 production. The IC50 of indomethacin determined using HTRF technology (1.0 ± 0.4 nM) was in agreement with previously published data (Palomer A. et al. 3 ± 2 nM ; Chan C.C. et al. 7 ± 1 nM).

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Prostaglandin E2 kit - 500 tests62P2APEG
-
Prostaglandin E2 kit - 10,000 tests62P2APEH
-

Companion products

DescriptionCat. noProduct insertMSDS
Diluent 2 - 20 ml62DL2DDD
Prostaglandin E2 kit standard62PG2CDA
-

ドキュメント