Estradiol ELISA kit

Name: Estradiol ELISA kit
SKU: DEIA-XY54
Rating: 5 (1 reviews)

Regulatory status: For research use only, not for use in diagnostic procedures.

COA

SDS

Datasheet

Summary

Sample

Saliva

Species Reactivity

Human

Intended Use

This Kit is an enzyme immunoassay for measurement of active free Estradiol, an estrogenic steroid, in saliva.

Contents of Kit

1. ##Microtiterwells##, 12 x 8 (break apart) strips, 96 wells;
2. ##Standard (Standard 0-5)##, 6 vials, 1 mL each, ready to use;
3. ##Control Low & High##, 2 vials, 1.0 mL each, ready to use;
4. ##Enzyme Conjugate##, 1 vial, 26 mL, ready to use;
5. ##Substrate Solution##, 1 vial, 25 mL, ready to use;
6. ##Stop Solution##, 1 vial, 14 mL, ready to use;
7. ##Wash Solution##, 1 vial, 30 mL;

Storage

2°C to 8°C

Performance Characteristics

##Assay Dynamic Range##
The range of the assay is between 0.8 - 93 pg/mL. Specificity The following materials have been evaluated for cross reactivity. The percentage indicates cross reactivity at 50% displacement compared to Estradiol.

##Sensitivity##
The lowest detectable level of Estradiol that can be distinguished from the Zero Standard is 0.4 pg/mL at the 95 % confidence limit.

##Reproducibility##
##Intra Assay##
The intra-assay variation was determined by 20 replicate measurements of 4 saliva samples using this ELISA kit.
The within assay variability is shown below:

##Inter Assay##
The inter-assay (between-run) variation was determined by duplicate measurements of five saliva samples over 10 different days runs.
The between assay variability is shown below:

##Inter-Lot##
The Inter-Lot (between-lot) variation was determined by triplicate measurements of five saliva samples in three different kit lots.
The between lot variability is shown below:

##Recovery##
Recovery of the CD ELISA was determined by adding increasing amounts of the analyte to three different saliva samples containing different amounts of endogenous analyte. Each sample (non-spiked and spiked) was assayed and analyte concentrations of the samples were calculated from the standard curve. The percentage recoveries were determined by comparing expected and measured values of the samples.

##Linearity##
Four samples (saliva) containing different amounts of analyte were serially diluted up to 1:128 with 0.9% NaCl and assayed with the CD ELISA. An assay linearity of 0.8 - 93 pg/mL has been identified as the usable range. Samples above this range must be diluted and re-run.

General Description

Estradiol (1,3,5(10)-estratriene-3,17β-diol; 17β-estradiol; E21) is a C18 steroid hormone with a phenolic A ring. This steroid hormone has a molecular weight of 272.4. It is the most potent natural Estrogen, produced mainly by the Graffian follicle of the female ovary and the placenta, and in smaller amounts by the adrenals, and the male testes. Estradiol (E2) is secreted into the blood stream where 98% of it circulates bound to sex hormone binding globulin (SHBG) and to a lesser extent to other serum proteins such as albumin. Only a small fraction circulates as free hormone or in the conjugated form. Estrogenic activity is effected via estradiol-receptor complexes which trigger the appropriate response at the nuclear level in the target sites. These sites include the follicles, uterus, breast, vagine, urethra, hypothalamus, pituitary and to a lesser extent the liver and skin. In non-pregnant women with normal menstrual cycles, estradiol secretion follows a cyclic, biphasic pattern with the highest concentration found immediately prior to ovulation. The rising estradiol concentration is understood to exert a positive feedback influence at the level of the pituitary where it influences the secretion of the gonadotropins, follicle stimulating hormone (FSH), and luteinizing hormone (LH), which are essential for follicular maturation and ovulation, respectively. Following ovulation, estradiol levels fall rapidly until the luteal cells become active resulting in a secondary gentle rise and plateau of estradiol in the luteal phase. During pregnancy, maternal serum Estradiol levels increase considerably, to well above the pre-ovulatory peak levels and high levels are sustained throughout pregnancy.

Standard Curve

##Standard## ##Absorbance Units##
Standard 0 (0 pg/mL) 2.08
Standard 1 (1 pg/mL) 1.90
Standard 2 (5 pg/mL) 1.62
Standard 3 (10 pg/mL) 1.40
Standard 4 (50 pg/mL) 0.58
Standard 5 (100 pg/mL) 0.21

Reconstitution And Storage

When stored at 2°C to 8°C unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date. Opened reagents must be stored at 2°C to 8°C. Microtiter wells must be stored at 2°C to 8°C. Once the foil bag has been opened, care should be taken to close it tightly again. Opened kits retain activity for two months if stored as described above.

Citations

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Documents

pdf COA

pdf SDS

pdf Datasheet

QA & Reviews

Customer Reviews

References

Late Follicular Phase Progesterone Elevation During Ovarian Stimulation Is Not Associated With Decreased Implantation of Chromosomally Screened Embryos in Thaw Cycles

OBSTETRICAL & GYNECOLOGICAL SURVEY

Authors: Hernandez-Nieto, Carlos; Lee, Joseph A.; Alkon-Meadows, Tamar; Luna-Rojas, Martha; Mukherjee, Tanmoy; Copperman, Alan B.; Sandler, Benjamin

Abstract

Although the physiologic mechanism remains unclear, late follicular phase progesterone elevation (LFPE) commonly occurs during controlled ovarian hyperstimulation (COH) for assisted-reproductive technology (ART) such as in vitro fertilization (IVF). Late follicular phase progesterone elevation has been associated with patient body mass index (BMI), ethnicity, ovarian reserve metrics, ovarian response to exogenous gonadotropins, number of follicles recruited during COH, dosage/ duration of stimulation cycles, and other factors. Despite the plethora of research on ART outcomes and ovarian stimulation, there is not definitive evidence of an effect of LFPE on embryonic quality or embryo reproductive potential. This retrospective analysis aimed to evaluate the impact of LFPE during COH on embryonic quality and aneuploidy, and analyze the pregnancy rates and IVF outcomes after the transfer of these embryos in a frozen euploid embryo transfer cycle. Patients diagnosed with infertility from a single academic ART center that underwent IVF/preimplantation genetic testing for aneuploidy (PGT-A) between September 2016 and March 2020 were included. The occurrence of LFPE was defined as serum progesterone level of 2.0 ng/mL or greater on the day of ovulation trigger. All patients, regardless of LFPE occurrence, underwent COH, intracytoplasmic sperm injection (ICSI), extended embryo culture, trophectoderm (TE) biopsy, and PGT-A. Patients were stratified into 2 groups based on progesterone levels: normal group (P = 2.0 ng/mL) and LFPE (P = 2.0 ng/mL). A subanalysis analyzed outcomes of single euploid FET cycles based on progesterone levels on the day of ovulation trigger included patients that completed IVF/PGT-A followed by synthetic endometrial preparation for a euploid FET cycle within the same date range as the primary analysis. Primary analysis outcomes included blastulation rate (viable blastocysts over total number of fertilized oocytes), utilizable blastocyst rate (blastocysts available for TE biopsy and vitrification), and ploidy rate. Embryonic quality was assessed, and embryos were classified into 3 cohorts based on morphological grading (good, moderate, fair). A total of 5141 cases were included in this analysis, with 4925 having normal progesterone and 216 having LFPE occur. In unadjusted analysis, significant differences between these 2 groups were found in patient age, BMI, serum progesterone, days of gonadotropins used, day of ovulation trigger, progesterone at trigger, estradiol at trigger, AMH, baseline FSH, AFC, and number of retrieved oocytes among cohorts. Oocyte maturation rate, aneuploidy rate, ferilization rate, percentage of good/ moderate/fair embryos, and blastulation rate were comparable between the normal and LFPE groups. Utilizable blastocyst rate was 65.3% (n = 22,654) in the normal progesterone cohort and 62.4% (n = 1337) in the LFPE cohort (P = 0.19). Adjustment for age, BMI, AMH, baseline FSH, days of stimulation, and oocytes retriever per case did not result in any statistically significant associations of LFPE with embryo aneuploidy (aOR, 1.04; 95% confidence interval, 0.86-1.27; P = 0.62) or odds of embryos being reported as inconclusive (aOR, 1.12; 95% confidence interval, 0.69-1.84; P = 0.62). A total of 5806 euploid FETs were included in the subanalysis, with 5617 cycles having patients with normal progesterone and remaining 189 cycles involving LFPE. Of the live births (n = 2250; normal n = 2198, LFPE n = 52), clinical and gestational outcomes were comparable between the 2 groups. The findings of this analysis indicate that LFPE defined as a serum progesterone value of 2.0 ng/mL drawn on the day of ovulation trigger is not associated with adverse pregnancy outcomes or an obstacle to embryo implantation potential.

Sex Steroids Induce Membrane Stress Responses and Virulence Properties in Pseudomonas aeruginosa

MBIO

Authors: Vidaillac, Celine; Yong, Valerie Fei Lee; Aschtgen, Marie-Stephanie; Qu, Jing; Yang, Shuowei; Xu, Guangfu; Seng, Zi Jing; Brown, Alexandra C.; Ali, Md Khadem; Jaggi, Tavleen K.; Sankaran, Jagadish; Foo, Yong Hwee; Righetti, Francesco; Nedumaran, Anu Maashaa; Mac Aogain, Micheal; Roizman, Dan; Richard, Jean-Alexandre; Rogers, Thomas R.; Toyofuku, Masanori; Luo, Dahai; Loh, Edmund; Wohland, Thorsten; Czarny, Bertrand; Horvat, Jay C.; Hansbro, Philip M.; Yang, Liang; Li, Liang; Normark, Staffan; Normark, Birgitta Henriques; Chotirmall, Sanjay H.

Abstract

Estrogen, a major female sex steroid hormone, has been shown to promote the selection of mucoid Pseudomonas aeruginosa in the airways of patients with chronic respiratory diseases, including cystic fibrosis. This results in long-term persistence, poorer clinical outcomes, and limited therapeutic options. In this study, we demonstrate that at physiological concentrations, sex steroids, including testosterone and estriol, induce membrane stress responses in P. aeruginosa. This is characterized by increased virulence and consequent inflammation and release of proinflammatory outer membrane vesicles promoting in vivo persistence of the bacteria. The steroid-induced P. aeruginosa response correlates with the molecular polarity of the hormones and membrane fluidic properties of the bacteria. This novel mechanism of interaction between sex steroids and P. aeruginosa explicates the reported increased disease severity observed in females with cystic fibrosis and provides evidence for the therapeutic potential of the modulation of sex steroids to achieve better clinical outcomes in patients with hormone-responsive strains. IMPORTANCE Molecular mechanisms by which sex steroids interact with P. aeruginosa to modulate its virulence have yet to be reported. Our work provides the first characterization of a steroid-induced membrane stress mechanism promoting P. aeruginosa virulence, which includes the release of proinflammatory outer membrane vesicles, resulting in inflammation, host tissue damage, and reduced bacterial clearance. We further demonstrate that at nanomolar (physiological) concentrations, male and female sex steroids promote virulence in clinical strains of P. aeruginosa based on their dynamic membrane fluidic properties. This work provides, for the first-time, mechanistic insight to better understand and predict the P. aeruginosa related response to sex steroids and explain the interindividual patient variability observed in respiratory diseases such as cystic fibrosis that are complicated by gender differences and chronic P. aeruginosa infection.