Estradiol Assay Kit

There is substantial research indicating that breast cancer tissues contain the enzymes necessary to synthesize estradiol (17β-estradiol), a steroid hormone which plays an important role in this disease. Two principal pathways are involved in the formation of estradiol in breast cancer tissues: the aromatase pathway, which transforms androgens into estrogens, and the sulfatase pathway, which converts estrone into estradiol via the 17β- HSD1 enzyme. Based on HTRF® technology, the Estradiol kit is a rapid and highly sensitive assay for measuring estradiol in whole cells, cell supernatants or biochemical enzyme assays.

Assay principle

The HTRF Estradiol assay can be run using purified 17β-HSD1 enzyme or live cells. The assay is a monoclonal antibody-based competitive assay that can be run in two steps: cell stimulation or enzyme incubation, followed by estradiol detection using Estradiol-XL665 and anti-Estradiol-Cryptate MAb. The protocol requires only a single two hour incubation following cell/enzyme stimulation.

Product specifications

Detection limit: 0.02 ng/mL
Dynamic range: 0.02 to 20 ng/mL
EC50: 0.8 ng/mL
S/B: 14
Specificity: Estrone 0.4%
Estriol 0.3%
Estrone 3 sulfate 0.002%
Testosterone 0.2%
Progesterone 0.002%.

Estradiol standard curve performed using protocol outlined in product insert.

Product performances

Detecting 17β-HSD1 activity

A human breast cancer cell-line, T-47D, was tested for the production of estradiol when stimulated with estrone using the HTRF Estradiol assay. Assay reagents were added directly to the cell culture plate following cell stimulation. As figure 2 shows, estradiol was detected using as few as 780 cells. The ability to miniaturize and run the assay in a single plate while not sacrificing assay performance makes HTRF Estradiol an ideal tool for screening. Published data (Palomer A. et al. 3 ± 2 nM ; Chan C.C. et al. 7 ± 1 nM).

T-47D cells were dispensed as indicated into 96-half-well plates. 50 µl MEM containing 10 nM Estrone was added and cells incubated for 20 hours. Estradiol assay reagents were added directly to cultured cells. Following a 2 hour incubation, fluorescence signals were read and estradiol concentrations calculated using a standard curve.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Estradiol kit - 500 tests62ESTPEG
Estradiol kit - 10,000 tests62ESTPEH

Companion products

DescriptionCat. noProduct insertMSDS
Diluent 2 - 20 ml62DL2DDD
Estradiol kit standard62ESTCDA