Estradiol Assay
There is substantial research that indicates 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. Cisbio Bioassays's HTRF® Estradiol assay is a rapid and highly sensitive assay for measuring estradiol in whole cells, cell supernatants or biochemical enzyme assays.
- Highly specific assay
- Simple protocol
- Live cell, supernatant, and biochemical-based protocols
- Serum compatible
- Amenable to HTS
- Miniaturization down to <10 µL
- Estradiol measurement
- Monitoring of 17β-HSD1 activity
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: | 0.019 ng/mL |
| Dynamic range: | 0.019 to 20 ng/mL |
| EC50: | 0.8ng/mL |
| S/B: | 14 |
| Specificity: |
Estrone 0.4% Estriol 0.3% Estrone 3 sulfate 0.002% Testosterone 0.2% Progesterone 0.002%. |
Figure 1: Estradiol standard curve performed using protocol outlined in product insert.
Assay Performance:
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 the HTRF Estradiol an ideal
tool for screening.
Figure 2: 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.
Ordering Info:
Estradiol Assay Reagents:
| Description | Quantity* | Cat no. | Product Insert | MSDS | ||
| Estradiol | 1,000 tests | 62ESTPEB | Request MSDS | |||
| Estradiol bulk** | 20,000 tests | 62ESTPEC | Request MSDS | |||
| *based on 20 µl assay volume **upon request |
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For more information or To Place an Order:
Europe and other countries
+33(0)466 796 705
