LBIS™ Mouse 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 |
※Check availability in the US with the distributor.
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Kit component
96 tests
| Antibody-coated Plate | 96 wells/1 plate |
|---|---|
| Mouse IL-6 Standard | 2 bottles |
| Buffer | 30 mL/1 bottle |
| Biotin-conjugated Antibody Solution | 100 μL/1 bottle |
| Peroxidase-conjugated Streptavidin Solution | 100 μL/1 bottle |
| TMB Solution | 12 mL/1 bottle |
| Buffer for Standard and Sample | 30 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™ Mouse IL-6 ELISA Kit is a sandwich ELISA kit for the quantitative measurement of mouse 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 | 2.05 - 500 pg/mL |
| Analysis sample | Mouse Serum/Plasma (EDTA)/Culture medium (D-MEM etc.)* |
| Sample amount | 25 μL (2x dilution) |
| Measurement duration | Approx 3 hours and 50 minutes |
| Wavelength | Primary wavelength : 450 nm Reference wavelength : 620 nm |
- It is tested by using the samples that standard substances have been added to the culture medium.
Measurement availability depends on the culture medium.
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 mouse serum samples at two concentrations.
| n＼ID | ID1 (pg/mL) | ID2 (pg/mL) |
|---|---|---|
| 1 | 281 | 18.7 |
| 2 | 285 | 19.1 |
| 3 | 281 | 18.7 |
| 4 | 282 | 18.7 |
| 5 | 291 | 19.6 |
| mean | 284 | 19.0 |
| SD | 4.24 | 0.40 |
| CV(%) | 1.5 | 2.1 |
- [Result]
- The CV (%) of mouse serum was 1.5-2.1%, indicating good repeatability.
Reproducibility (between run precision)
Reproducibility was conducted by measuring mouse serum samples at three concentrations each for four days.
| Day＼ID | ID3 (pg/mL) | ID4 (pg/mL) | ID5 (pg/mL) |
|---|---|---|---|
| 1 | 246 | 84.0 | 15.1 |
| 2 | 230 | 78.8 | 13.9 |
| 3 | 254 | 86.2 | 15.3 |
| 4 | 244 | 84.9 | 14.4 |
| mean | 244 | 83.5 | 14.7 |
| SD | 10.0 | 3.24 | 0.645 |
| CV(%) | 4.1 | 3.9 | 4.4 |
- [Result]
- The CV (%) of mouse serum was 3.9-4.4%, indicating good reproducibility.
Dilution Linearity Test
Mouse serum and plasma (EDTA), as well as culture medium (D-MEM and 10% FCS/D-MEM), were serially diluted twofold using the provided buffer and measured in duplicate to evaluate linearity.
- [Result]
- All measurement results showed good linearity within the measurement range.
Spike and Recovery Test
Standard solutions at three concentrations were added to mouse serum (EDTA), as well as culture medium (D-MEM and 10% FCS/D-MEM), then spike recovery tests were conducted, with each measurement performed in duplicate.
Mouse serum
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 4.8 | - | - |
| 22.8 | 26.9 | 22.1 | 96.9 |
| 46.0 | 49.0 | 44.2 | 96.0 |
| 96.4 | 95 | 90.1 | 93.4 |
| 197.0 | 193 | 188.2 | 95.5 |
| mean | 95.5 | ||
Mouse plasma (EDTA)
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 4.4 | - | - |
| 28.1 | 30.6 | 26.2 | 93.0 |
| 56.0 | 55.0 | 50.5 | 90.2 |
| 115.6 | 107 | 102.1 | 88.4 |
| 227.3 | 206 | 201.8 | 88.8 |
| mean | 90.1 | ||
Culture medium (D-MEM)
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 0.0 | - | - |
| 23.0 | 24.1 | 24.1 | 105.0 |
| 48.3 | 47.7 | 47.7 | 98.7 |
| 97.5 | 93 | 93.0 | 95.3 |
| 194.4 | 205 | 205.1 | 105.5 |
| mean | 101.1 | ||
Culture Medium (10% FCS/D-MEM)
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 0.0 | - | - |
| 28.1 | 27.6 | 27.6 | 98.1 |
| 56.0 | 60.2 | 60.2 | 107.5 |
| 115.6 | 121 | 120.7 | 104.4 |
| 227.3 | 237 | 237.2 | 104.3 |
| mean | 103.6 | ||
- [Result]
- Good recovery rates were confirmed.
Cross-reactivity with similar proteins
Proteins similar to mouse IL-6, including IL-6 of other species and other interleukins, were tested to determine if they could be detected.
| Proteins | Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Cross-reactivity (%) |
|---|---|---|---|
| Human IL-6 | 500 | 0.034*1 | 0.01 |
| Rat IL-6 | 500 | 0.214*1 | 0.04 |
| Mouse IL-4 | 1,000 | 0.01*1 | 0.001 |
| Mouse IL-7 | 1,000 | -*2 | - |
| Mouse gp130 | 1,000 | -*2 | - |
| Mouse IL-6Rα | 1,000 | -*2 | - |
*1 Values outside the quantification range; shown as reference
*2 -: Below the lower limit of quantification
- [Result]
- Very low reactivity with similar proteins was confirmed.
Correlation with conventional product
For mouse serum samples, measurements were conducted using this kit and competitor Company A. The measurement values obtained using both products were then compared.
- [Result]
- Good correlation was confirmed.
Detection Performance Comparison
Measurements were conducted on mouse serum samples using this kit and competitor Company A to compare their detectability.
| ID | Fujifilm Wako (pg/mL) |
Company A (pg/mL) |
|---|---|---|
| 1 | 1.71 | Not detectable |
| 2 | 6.50 | 5.52 |
| 3 | 6.39 | 9.02 |
| 4 | 7.01 | 7.67 |
| 5 | 9.67 | 7.53 |
| Proportion of samples successfully measured |
4/5 samples | 1/5 samples |
Values within the calibration range are indicated in red; samples outside the
range are shown as reference values.
- [Result]
- This kit demonstrated high detection sensitivity, successfully detecting four out of five samples within the calibration curve range.
FAQ
About samples
- What anticoagulants should I use?
- We recommend using EDTA 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 ultipette 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 mouse IL-6 standard?
- The amount of purified water to be added varies by lot. Please reconstitute the mouse 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℃.
Mouse 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.
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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