Estradiol assay

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.

Features

Highly specific assay
Simple protocol
Live cell, supernatant, and biochemical-based protocols
Serum compatible
Amenable to HTS
Miniaturization down to <10 µL

Applications

Estradiol measurement
Monitoring of 17β-HSD1 activity

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.

Detection limit:	0.019 ng/mL
Dynamic range:	0.019 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

Estradiol standard curve performed using protocol outlined in product insert.

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).

Estradiol standard curve

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

Description	Cat. no	Product insert	MSDS
Estradiol kit - 1,000 tests	62ESTPEB	pdf	pdf
Estradiol kit - 20,000 tests	62ESTPEC	pdf	pdf

Companion products

Description	Cat. no	Product insert	MSDS
Diluent 2 - 20 ml	62DL2DDD	-	pdf
Reconstitution buffer 2 - 13 ml	62RB2RDD	-	-

Estradiol assay

Features

Applications

Related applications and services

Assay Principle

Product Specifications

Product Performances

Detecting 17β-HSD1 activity:

Part#, inserts & MSDS

Ordering Info

Companion products

Literature

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