0 item(s)
View cart
You have no items in your shopping cart.
Product Description
A sandwich-format solid-phase enzyme-linked immunosorbent assay (ELISA) for quantitative determination of natural or recombinant human Interleukin-6 (hIL-6) in biological samples. Wells are pre-coated with a mouse anti-hIL-6 capture antibody. Samples and calibration standards are incubated in the wells, followed by a biotinylated anti-hIL-6 detection antibody and streptavidin-HRP (SA-HRP). TMB substrate is converted by HRP to a blue product (max absorbance ~655 nm) and then stopped to yield a yellow product read at 450 nm (with 620–650 nm reference). hIL-6 concentrations are calculated by fitting a 4-parameter logistic (4-PL) calibration curve from supplied standards.
Technical Specifications
| Parameter | Details |
|---|---|
| Package size | 96 tests/Kit |
| Assay format | Double-antibody sandwich ELISA (solid-phase) |
| Sample volume per well | 100 µL (standards, controls and samples) |
| Linearity & Range | 2–512 pg/mL, R² ≥ 0.990 |
| Lower Limit of Quantification (LLOQ) | 2 pg/mL |
| Specificity / Cross-reactivity | No cross-reactivity with IL-2, IL-10, IL-12, IFN-γ, IFN-β, IL-6Rα, Rat IL-6 or Mouse IL-6 |
| Typical calibration curve model | 4-parameter logistic (4-PL) regression; example parameters A=7.86930, B=-1.04651, C=760.10625, D=0.00086, R²=0.99999 |
| Calibration standards (pg/mL) | 512, 256, 128, 32, 8, 2, 1, 0 (NCS) |
| Incubation times | Sample incubation 1 hour at RT (600 rpm recommended); Biotinylated antibody 45 minutes at RT; Streptavidin-HRP 30 minutes at RT; TMB substrate 10 minutes at RT |
| Washing | Wash plates with 300 µL 1×Wash Buffer; typical wash cycles: after sample incubation 3×, after biotin 3×, before TMB 5× (as described) |
| Detection / Readout | Optical density read at 450 nm, with reference/correction wavelength 620–650 nm; read within 10 minutes after stop |
| Reagent concentrations / dilutions | Anti-hIL-6: Biotinylated Conjugate (200×) — dilute 200× with Diluent prior to use; Streptavidin-HRP (100×) — dilute 100× with Diluent prior to use; Wash Buffer Concentrate (10×) — dilute 10× |
| Storage of key components (opened) | Anti-hIL-6 Microtiter Strips: store in bag with desiccant at 2–8 °C up to 90 days; Reconstituted Calibration Standard: aliquot and store ≤-18 °C; other components: follow label instructions |
Features
- High sensitivity: LLOQ 2 pg/mL and linear range down to 2 pg/mL enabling detection of low-level hIL-6.
- High specificity: No observed cross-reactivity with related cytokines and IL-6 from rat or mouse.
- Ready-to-use coated strips: Microtiter strips pre-coated with capture antibody in sealed bag with desiccant for convenience and reduced setup time.
- Complete kit with standards and reagents: Includes lyophilized calibration standard, reconstitution solution, diluent, conjugates, SA-HRP, substrate and stop solution for a full quantitative assay.
- Standardized quantitation: Supplied calibration series and 4-PL fitting enable robust quantification of hIL-6.
Applications
- Quantitation of human IL-6 in serum
- Quantitation of human IL-6 in cell culture supernatants
- In-process samples, harvested bulk, drug substance and drug product testing (research applications)
- Research use in immunology, cell biology and bioprocess monitoring
Kit Contents
Anti-hIL-6 Microtiter Strips (pre-coated)Sealed in bag with desiccant, store at 2-8 °C (opened strips: up to 90 days)
hIL-6 Calibration Standard (lyophilized)Lyophilized; reconstitute with 500 µL Reconstitution Solution; aliquot and store ≤-18 °C, avoid freeze-thaw
Reconstitution Solution2-8 °C (follow label)
Diluent2-8 °C (follow label)
Wash Buffer Concentrate (10×)2-8 °C; dilute 1:10 with ultrapure water before use
Anti-hIL-6: Biotinylated Conjugate (200×)2-8 °C; centrifuge before use; dilute 200× with Diluent and prepare fresh
Streptavidin-HRP (100×)2-8 °C; centrifuge before use; dilute 100× with Diluent and prepare fresh
TMB SubstrateSealed, protect from light; equilibrate to RT before use
Stop Solution (1 M HCl)2-8 °C; corrosive—avoid contact with eyes, skin, clothing
Sealing FilmRoom temperature (RT)
* Total: 96 tests/Kit | Method: ELISA (sandwich)
Attention
• For Research Use Only (not for clinical or diagnostic use).
• Qualified personnel only; validate assay in-house before routine use.
• Do not mix reagents from different lot numbers.
• Reconstituted Calibration Standard, 1× Biotinylated Conjugate and 1× Streptavidin-HRP are recommended single-use; prepare freshly.
• Use fresh ultrapure/sterile water; if concentrates are cloudy, heat to 37 °C until clear.
• Avoid drying wells before substrate incubation and avoid bubbles during pipetting.
• Stop Solution is 1 M HCl — corrosive; avoid contact with eyes, skin and clothing.
• Avoid samples containing sodium azide (NaN3) as it will inactivate HRP and lead to underestimation.
• TMB Substrate should be colorless; discard if discolored.
• Use sterile disposable consumables and avoid cross-contamination; recommended to wipe benches and equipment with 75% ethanol.
Quality Management & Certifications
Quality System
Download QMSQMS (ISO, GMP)
Download CertificateQuality Advantages
View ReportQuality Control Process
Download ProcessTechnical Resources & Downloads
Frequently Asked Questions (FAQ)
Q1: What sample types are compatible with this kit?
This kit is validated for human biological samples such as serum and cell culture supernatants. It can also be used for in-process samples, harvested bulk, drug substance and drug product for research purposes. Samples should be clear and insoluble material removed by centrifugation or filtration; appropriate dilutions may be required.
Q2: What is the assay sensitivity and working range?
The assay LLOQ is 2 pg/mL with a linear quantitative range of 2–512 pg/mL (R² ≥ 0.990).
Q3: How should I prepare the calibration standards?
Reconstitute the lyophilized calibration standard with 500 µL Reconstitution Solution. Prepare serial dilutions as described to obtain standards at 512, 256, 128, 32, 8, 2, 1 and 0 pg/mL. Follow the exact volumes and dilution scheme in the manual.
Q4: How should I store opened kit components?
Store the kit at 2–8 °C. Keep unused microtiter strips sealed with desiccant at 2–8 °C (opened strips stable up to 90 days). Aliquot reconstituted calibration standard and store at ≤-18 °C; avoid freeze-thaw cycles. Prepare 1× conjugates fresh and use single-use where recommended.
Research Use Only
✔
For Research Use Only (RUO) — Not for clinical or diagnostic use – This product is intended for laboratory research purposes only and not for clinical or regulatory diagnostic use.
✔
Not intended for use in USDA or FDA regulated diagnostic testing or official compliance testing.
When can I expect my order to ship?
Most orders are filled and shipped within 2-3 business days from the time they are received.
Our standard shipping usually take 2-5 days.
We also provide express shippping for time-sensitive deliveries.
Email contact@biofargo.com if you have any requirements.
