Key Features
- Specific Detection of Estrone-3-Sulfate: Designed to measure the sulfated estrogen metabolite E3S. Cross-reactivity with free estrogens (E1, E2, E3) is <0.1%.
- Transparent Cross-Reactivity Data: DHEA-S ≤20%; other estrogen sulfates (E2-3-sulfate, E2-17β-sulfate) ≤7%. Full details provided to support experimental design.
- High Sensitivity & Broad Range: Minimum detectable concentration <1.0 pg/mL; quantitative range 1.0–400 pg/mL, supported by a six-point standard curve (0, 25, 50, 100, 200, 400 pg/mL).
- Built-in 5× Sample Dilution: Protocol automatically dilutes samples 5-fold (10 µL sample + 40 µL diluent). Multiply the interpolated concentration by 5 to obtain the actual sample value.
- Recombinant Standard Protein: Standards are produced in a eukaryotic expression system, ensuring proper folding and immunoreactivity.
- Validated Sample Types: Human serum, EDTA/citrate/heparin plasma, cell culture supernatant, and tissue homogenates.
- One-Step Sandwich ELISA: Simultaneous incubation of sample/standard and HRP-detection antibody; total hands-on time approximately 2 hours.
- Reliable Performance: Intra- and inter-assay CV <15%; standard curve linearity R ≥0.9900.
- Removable Strips & Scalable Formats: 48-well and 96-well configurations; up to 88 samples per plate (single wells).
- Storage & Stability: 6-month shelf life at 2–8°C, protected from light and moisture; cold-chain shipping.
Product Description
Estrone-3-sulfate (E3S) is a major circulating sulfated estrogen and a key reservoir for biologically active estrogens in hormone-dependent tissues. It serves as a biomarker and metabolic intermediate in studies of steroidogenesis, endocrine disruption, pregnancy physiology, and hormone-responsive cancers. Accurate quantitation of E3S, independent of free estrogens, is essential for delineating sulfatase and sulfotransferase pathways in human research.
The Yanda-HUMAN-E3S ELISA Kit employs a one-step double-antibody sandwich ELISA method. A capture antibody pre-coated on the microplate recognizes the 3-sulfate moiety and the steroidal backbone of E3S. Samples and standards are co-incubated with an HRP-conjugated detection antibody. Following a wash step, TMB substrate generates a signal proportional to the E3S concentration, and absorbance is read at 450 nm.
Sample Collection & Preparation
| Sample Type | Procedure |
|---|---|
| Serum | Collect in pyrogen/endotoxin-free tubes without anticoagulant. Let blood clot, then centrifuge at 3000 rpm for 10 min. Separate serum promptly. |
| Plasma | Use EDTA, citrate, or heparin. Centrifuge at 3000 rpm for 30 min; collect supernatant. |
| Cell Culture Supernatant | Centrifuge at 3000 rpm for 10 min to remove cells and debris. |
| Tissue Homogenate | Homogenize tissue in ice-cold normal saline; centrifuge at 3000 rpm for 10 min. Use supernatant. |
Storage: If not tested immediately, aliquot and store at -20°C. Avoid repeated freeze-thaw cycles. Thaw completely at room temperature and mix gently.
⚠️ Critical Dilution Factor:
The assay protocol dilutes samples 5-fold in the well (10 µL sample + 40 µL Sample Diluent). The concentration read from the standard curve is the diluted value. Multiply the result by 5 to obtain the concentration in the original specimen.
For samples with unknown E3S levels, we recommend performing a pilot experiment to determine if additional pre-dilution is necessary.
Technical Specifications
| Parameter | Specification |
|---|---|
| Standard Concentrations (S0–S5) | 0, 25, 50, 100, 200, 400 pg/mL |
| Quantitative Range | 1.0 – 400 pg/mL |
| Analytical Sensitivity | < 1.0 pg/mL |
| Accuracy (Linearity) | Standard curve R ≥ 0.9900 |
| Precision | Intra-assay CV < 15%; Inter-assay CV < 15% |
| Specificity | Specific for estrone-3-sulfate. See cross-reactivity table below. |
| Sample Dilution Factor | 5× (automatically performed in the well) |
| Shelf Life | 6 months |
| Storage & Transport | 2–8°C, protected from light and moisture; cold-chain shipping |
Cross-Reactivity Profile
| Compound | Cross-Reactivity |
|---|---|
| DHEA-S (dehydroepiandrosterone sulfate) | ≤ 20% (significant; consider when DHEA-S levels are elevated) |
| E2-3-sulfate / E2-17β-sulfate | ≤ 7% |
| Free Estrone (E1) | < 0.1% |
| Free Estradiol (E2) | < 0.1% |
| Free Estriol (E3) | < 0.1% |
| Other soluble structural analogues | No cross-reactivity observed |
Representative Standard Curve
Representative performance data, including spike-recovery and dilution linearity, are available in the product instruction manual. For additional validation details, please contact technical support.
Kit Components
96-Well Format
| Component | Quantity | Notes |
|---|---|---|
| Microelisa Stripplate | 12 strips × 8 wells | Pre-coated with anti-E3S antibody |
| Standard (0.3 mL/vial) | 6 vials | Ready-to-use liquid, 0–400 pg/mL |
| Sample Diluent | 6 mL | – |
| HRP-Conjugated Detection Antibody | 10 mL | Ready-to-use |
| 20× Wash Buffer | 25 mL | Dilute to 1× before use |
| Substrate A | 6 mL | – |
| Substrate B | 6 mL | – |
| Stop Solution | 6 mL | – |
| Plate Sealers | 2 sheets | – |
| Instruction Manual | 1 copy | – |
| Self-sealing Bag | 1 piece | For unused strip storage |
48-Well Format
| Component | Quantity | Notes |
|---|---|---|
| Microelisa Stripplate | 12 strips × 4 wells | Pre-coated, removable |
| Standard (0.3 mL/vial) | 6 vials | Ready-to-use liquid |
| Sample Diluent | 3 mL | – |
| HRP-Conjugated Detection Antibody | 5 mL | Ready-to-use |
| 20× Wash Buffer | 15 mL | Dilute to 1× |
| Substrate A | 3 mL | – |
| Substrate B | 3 mL | – |
| Stop Solution | 3 mL | – |
| Plate Sealers | 2 sheets | – |
| Instruction Manual | 1 copy | – |
| Self-sealing Bag | 1 piece | – |
Note: The 20× Wash Buffer may crystallize during cold storage; warm in a water bath until crystals dissolve completely before dilution.
Assay Procedure Summary
- Equilibrate: Allow the sealed foil pouch to reach room temperature for 20 min. Remove required strips; seal unused strips and store at 4°C.
- Add Standards & Samples:
- Standard wells: 50 µL of each standard.
- Sample wells: 10 µL sample + 40 µL Sample Diluent.
- Blank well(s): Leave empty.
- Add Detection Antibody: Add 100 µL HRP-conjugated detection antibody to all wells except blank. Cover with plate sealer and incubate at 37°C for 60 minutes.
- Wash: Aspirate and blot. Fill each well with 1× Wash Buffer, let stand for 1 min, then discard. Repeat for 5 washes. An automated plate washer may be used (350 µL/well, 1-min soak).
- Substrate: Add 50 µL each of Substrate A and Substrate B. Incubate in the dark at 37°C for 15 minutes.
- Stop & Read: Add 50 µL Stop Solution. Measure absorbance at 450 nm within 15 minutes.
Total assay time: approximately 2 hours.
Materials Required but Not Supplied:
- Microplate reader (450 nm)
- Adjustable pipettes and tips (0.5–1000 µL)
- 37°C incubator or water bath
- Distilled/deionized water
- Absorbent paper, wash bottle or automated plate washer
Throughput
| Format | Max. Number of Samples (Single Wells) |
|---|---|
| 48-well | 39–40 samples |
| 96-well | 85–88 samples |
Includes 6 standard wells and 1–2 blank wells. For duplicate measurements, divide the sample number accordingly.
Expand Your Estrogen Research Panel
Pair this human E3S ELISA kit with the following assays for a comprehensive endocrine metabolism analysis:
- Human Estrone (E1) ELISA Kit – quantify the unconjugated parent estrogen that is released from E3S by steroid sulfatase.
- Human Estradiol (E2) ELISA Kit – measure the most potent natural estrogen to evaluate the endpoint of the sulfatase pathway.
- Human Apolipoprotein E (ApoE) ELISA Kit – Quantitative Detection for Research.
- Human Dihydrotestosterone (DHT) ELISA Kit – High‑Specificity Sandwich Assay for Androgen Research
Frequently Asked Questions
Q: Why must I multiply the result by 5?
A: The assay dilutes the sample 5-fold directly in the well (10 µL sample + 40 µL diluent). The standard curve is constructed under the same condition; therefore, the interpolated concentration reflects the diluted sample. Multiply by 5 to get the actual concentration in the original specimen.
Q: How significant is the DHEA-S cross-reactivity?
A: DHEA-S cross-reactivity is typically ≤20%. If your samples contain elevated DHEA-S (e.g., in certain endocrine disorders), this may contribute to the measured signal. We recommend assessing DHEA-S levels in parallel and discussing data interpretation with our technical support team.
Q: Can this kit be used to measure free estrone or estradiol?
A: No. The antibodies are specifically designed against the 3-sulfate group and steroidal backbone. Cross-reactivity with free E1, E2, and E3 is <0.1%, making the kit unsuitable for quantifying unconjugated estrogens.
Q: What if my sample reads above 400 pg/mL after applying the 5× factor?
A: Pre-dilute the sample further with the provided Sample Diluent (e.g., 1:2 or 1:5), re-assay, and multiply by the 5× in-well dilution plus the additional dilution factor.
Q: How should I handle tissue homogenates?
A: Homogenize tissue in ice-cold normal saline or PBS, centrifuge to clarify, and test the supernatant. Since matrix effects can vary, perform a pilot dilution series to determine the optimal pre-dilution factor if necessary.
Q: What is the source of the standards?
A: The standards are recombinant proteins produced in a eukaryotic cell expression system, which closely mimics the native conformation and immunoreactivity of human E3S.
Ordering & Contact Information
| Item | Details |
|---|---|
| Catalog Number | Yanda-HUMAN-E3S |
| Formats | 48-well / 96-well |
| Package Dimensions | 16 cm × 12.5 cm × 9.5 cm (neutral white box) |
| Shipping | Cold-chain transport (2–8°C), protected from light and moisture |
| Storage | 2–8°C; do not freeze |
| Shelf Life | 6 months |
| Intended Use | For research use only, not for diagnostic procedures |
Contact us
- WhatsApp: +86 198 3186 0669
- Email: songhawyble@gmail.com
For pricing, bulk orders, or customization inquiries, please reach out via the above channels.
Technical Documents & Support
- Instruction Manual – Available for download [insert link] or upon request.
- MSDS – Supplied with the kit or available on request.
- Technical Support – For detailed validation data, protocol assistance, or troubleshooting, contact us via WhatsApp or email.
Disclaimer
This product is for laboratory research use only and is not approved for clinical diagnosis, treatment, or prognosis. The manufacturer assumes no responsibility for consequences arising from improper use, protocol deviations, or unauthorized applications. Users are responsible for validating the assay in their own sample matrices and experimental conditions.
Why Choose Yanda Bio
- Unmatched diversity: Over 6,000 validated ELISA,targets across humans, rats, mice, rabbits, pigs, cattle, fish, microorganisms and insects Elisa kit 15+ species.
- Proven performance: Kits are highly reproducible, specific, and sensitive, with custom sensitivity options.
- Academic trust: Long‑term collaborations with 60+ universities in China.
- Free shipping: On any order of 3 kits.
- Complimentary elisa testing service: We run the assay, provide a video, and deliver all data at no cost.
- High‑sensitivity options: NHS‑biotin‑amplified kits available for ultra‑low detection.
- Immunogenicity testing: Dedicated immunogenicity elisa testing services for anti‑drug antibody evaluation.
Disclaimer: This kit is for research use only and is not intended for clinical diagnosis or therapeutic decision‑making. For clinical hormonal assessment, standard estradiol or estrone tests should be employed. This human E3S ELISA kit is validated for human samples only and must not be used with non‑human specimens. Follow the protocol exactly; any deviations are the responsibility of the experimenter. Yanda Bio shall not be liable for consequences arising from improper use.
Yanda Bio – Precision ELISA Kits, Expert elisa testing service, and immunogenicity elisa testing services for your endocrine pharmacology and oncology research.
