LBIS™ Human IL-6 ELISA Kit
- for Immunochemistry
- Manufacturer :
- FUJIFILM Wako Pure Chemical Corporation
- Storage Condition :
- Keep at 2-10 degrees C.
- GHS :
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96Tests
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In stock in Japan |
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Kit component
96 tests
| Antibody-coated Plate | 96 wells/1 plate |
|---|---|
| Human IL-6 Standard | 1 bottle |
| Buffer | 60 mL/1 bottle |
| Biotin-conjugated Antibody | 1 bottle |
| Peroxidase-conjugated Streptavidin Solution | 100 μL/1 bottle |
| TMB Solution | 12 mL/1 bottle |
| Stop Solution | 12 mL/1 bottle |
| Wash Solution (10x) | 100 mL/1 bottle |
| Plate Seal | 4 sheets |
Product Overview
Interleukin-6 (IL-6) is a pro-inflammatory cytokine that induces the differentiation of B cells into antibody-producing cells and plays a broad and important role in immune responses, inflammatory reactions, and antibody production. IL-6 is also implicated in the pathogenesis of autoimmune diseases such as rheumatoid arthritis, inflammatory conditions such as gout, and cytokine storms that cause acute respiratory distress syndrome (ARDS). Consequently, it is attracting considerable attention as a therapeutic target.
LBIS™ Human IL-6 ELISA Kit is a sandwich ELISA kit for the quantitative measurement of human IL-6. This kit enables highly sensitive quantification of IL-6 in biological samples. The standard included in the kit is a recombinant protein produced in Escherichia coli.
Kit Performance
| Assay target | IL-6 |
|---|---|
| Calibration curve range | 1.16 - 500 pg/mL |
| Analysis sample | Human Serum/Plasma (EDTA, heparin)/Cell culture supernatant* |
| Sample amount | 50 μL (2x dilution) |
| Measurement duration | Approx 3 hours and 50 minutes |
| Wavelength | Primary wavelength : 450 nm Reference wavelength : 620 nm |
When using cell culture supernatant as a sample, please make sure to perform recovery test and dilution linearity test on the culture medium before use.
Example of Calibration Curve
Data
Repeatability (within run precision)
Repeatability was conducted by measuring fivefold assays of human serum samples at two concentrations.
| n＼ID | ID1 (pg/mL) | ID2 (pg/mL) |
|---|---|---|
| 1 | 170 | 36.1 |
| 2 | 169 | 35.8 |
| 3 | 175 | 35.6 |
| 4 | 171 | 36.2 |
| 5 | 173 | 36.4 |
| mean | 172 | 36.0 |
| SD | 2.4 | 0.32 |
| CV(%) | 1.4 | 0.89 |
- [Result]
- The CV (%) of human serum was 0.89-1.4%, indicating good repeatability.
Reproducibility (between run precision)
Reproducibility was conducted by measuring human serum samples at three concentrations each for four days.
| Day＼ID | ID3 (pg/mL) | ID4 (pg/mL) | ID5 (pg/mL) |
|---|---|---|---|
| 1 | 186 | 73.9 | 20.7 |
| 2 | 207 | 80.6 | 23.1 |
| 3 | 184 | 71.9 | 20.8 |
| 4 | 192 | 78.5 | 22.9 |
| mean | 192 | 76.2 | 21.9 |
| SD | 10.4 | 4.0 | 1.3 |
| CV(%) | 5.4 | 5.3 | 6.0 |
- [Result]
- The CV (%) of human serum was 5.3-6.0%, indicating good reproducibility.
Dilution Linearity Test
Human serum and plasma (EDTA/heparin), as well as cell culture supernatant spiked with the standard, were serially diluted twofold using the provided buffer and measured in duplicate to evaluate linearity.
Human serum
Human plasma (EDTA)
Human plasma (heparin)
Human PBMC supernatants ± PHA stimulation (in RPMI-1640 medium with 10% FBS)
- Human PBMC supernatants stimulated with PHA (phytohemagglutinin, simulating IFN-γ secretion) were measured at 1:25 dilution.
- [Result]
- All measurement results showed good linearity within the measurement range.
Spike and Recovery Test
Standard solutions at three concentrations were added to human serum/plasma (EDTA), then spike recovery tests were conducted, with each measurement performed in duplicate.
Human serum
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 6.84 | - | - |
| 40 | 46.4 | 39.6 | 99 |
| 160 | 157 | 150 | 93.8 |
| 400 | 388 | 381 | 95.3 |
| mean | 96.0 | ||
Human plasma (EDTA)
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 33.9 | - | - |
| 40 | 75.1 | 41.2 | 103 |
| 160 | 193 | 159 | 99.4 |
| 400 | 443 | 409 | 102 |
| mean | 102 | ||
Human plasma (heparin)
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 38.5 | - | - |
| 40 | 79.8 | 41.3 | 103 |
| 160 | 196 | 158 | 98.8 |
| 400 | 435 | 397 | 99.3 |
| mean | 100 | ||
- [Result]
- Good recovery rates were confirmed.
Example of Sample Measurement
Human serum and plasma (EDTA/heparin) samples were measured and compared using this kit and the competitor.
Human serum
| ID＼n | Fujifilm Wako (pg/mL) | Company A (pg/mL) |
|---|---|---|
| 1 | 9.27 | N.D. |
| 2 | N.D. | N.D. |
| 3 | N.D. | N.D. |
| 4 | 4.74 | 5.42 |
| 5 | 7.95 | 4.12 |
| 6 | 3.50 | N.D. |
| 7 | 3.15 | N.D. |
| 8 | 125 | 141 |
| 9 | 0.862 | N.D. |
| 10 | 35.3 | 5.50 |
Human plasma (EDTA)
| ID＼n | Fujifilm Wako (pg/mL) | Company A (pg/mL) |
|---|---|---|
| 11 | 39.8 | 35.0 |
| 12 | 16.4 | 7.49 |
| 13 | 36.0 | 11.7 |
| 14 | 4.17 | N.D. |
| 15 | 2.46 | N.D. |
| 16 | 5.72 | N.D. |
| 17 | 4.31 | N.D. |
| 18 | 21.0 | 15.9 |
| 19 | 23.3 | 3.61 |
| 20 | 2.02 | N.D. |
Human plasma (heparin)
| ID＼n | Fujifilm Wako (pg/mL) | Company A (pg/mL) |
|---|---|---|
| 21 | 5.55 | N.D. |
| 22 | N.D. | N.D. |
| 23 | 4.89 | N.D. |
| 24 | 2.11 | N.D. |
| 25 | N.D. | N.D. |
| 26 | N.D. | N.D. |
| 27 | 3.74 | 5.46 |
| 28 | 3.93 | N.D. |
| 29 | 1.91 | N.D. |
| 30 | 2.96 | N.D. |
N.D.: Below the lower limit of quantification
Red: Values outside the quantification range; shown as reference
- [Result]
- This product was able to detect IL-6 even in samples that were below the lower limit of quantification of another manufacturer's kit.
FAQ
About samples
- What anticoagulants should I use?
- We recommend to use EDTA or heparin as anticoagulant.
- How should I store and prepare the samples?
- Samples should be immediately assayed or stored below -35℃ until assay. Before starting assay, shake thawed samples sufficiently. Do not repeat freeze-and-thaw cycles. Dilution of a sample should be made in a PP or PE test tube using the provided buffer prior to adding them to wells.
About kit usage
- What instruments, and equipment are required for the assay using this kit?
- The instruments and equipment required for the use of this kit are listed below;
- Deionized water (or distilled water)
- Test tubes for preparation of standard solution series
- Glassware for dilution of Wash Solution (10x) (a graduated cylinder, a bottle)
- Pipettes (disposable tip type). One should be able to deliver 50 μL precisely, and another for 100 – 1,000 μL
- Syringe-type repeating dispenser like Eppendorf multipette plus which can dispense 100 μL
- Paper towel to remove washing buffer remaining in wells
- A vortex-type mixer
- A shaker for 96 well-plate (600 – 1,200 rpm)
- An automatic washer for 96 well-plate (if available), or a wash bottle with a jet nozzle
- A 96 well-plate reader (450 ± 10 nm, 620 nm :600 - 650 nm)
- Software for data analysis
- How should I use the human IL-6 standard?
- The amount of purified water to be added varies by lot. Please reconstitute the human IL-6 standard with the amount of purified water specified in the “Reconstitution of standard” section on this product page to prepare the original standard solution (5 ng/mL). Then, dilute the solution using the kit-provided buffer that has been equilibrated to room temperature.
- What are the storage conditions for each reagent?
- [Buffer & TMB Solution]
Use only volume you need for your assay. Remaining reagents should be stored at 2-10℃ closing the cap tightly. Once opened, we recommend using as soon as possible to avoid influence by environmental condition.
[Stop Solution]
Close the cap tightly and store at 2-10℃.
[Human IL-6 Standard & Biotin-conjugated Antibody]
The reconstituted standard solution (original standard solution) should be used within 2 weeks if stored in a refrigerator. Dispose the remaining diluted standard solutions after use.
[Peroxidase-conjugated Streptavidin Solution]
Remaining working solution (already diluted) should be disposed. The rest of the undiluted solution (unused): close the cap tightly and store at 2-10℃. Once opened, we recommend using as soon as possible to avoid influence by environmental condition.
Prepare reagent solutions in appropriate volume for your assay.
Overview / Applications
Property
Manufacturer Information
Alias
- 635-42311 (Previous Code)
AKH-IL6 (Previous Code)
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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