LBIS (TM) Mouse IFN-γ 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 IFN-γ 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
Interferon-γ (IFN-γ) is an important cytokine involved in both innate and adaptive immunity. Cells known to produce IFN-γ include natural killer (NK) cells and T cells. Secreted IFN-γ induces a wide range of immune responses, including antiviral activity, activation of macrophages, maturation of dendritic cells, and induction of T-cell differentiation. IFN-γ also plays a significant role in antitumor immunity, with reported effects including suppression of metastasis, induction of apoptosis, and inhibition of angiogenesis.
LBIS™ Mouse IFN-γ ELISA Kit is a sandwich ELISA kit for the quantitative measurement of mouse IFN-γ. This kit enables highly sensitive quantification of IFN-γ in biological samples. The standard included in the kit is a recombinant protein produced in Escherichia coli.
Kit Performance
| Assay target | IFN-γ |
|---|---|
| Calibration curve range | 2.05 – 500 pg/mL |
| Analysis sample | Mouse Serum/Plasma (EDTA, heparin) |
| Sample amount | 25 μL (2x dilution) |
| Measurement duration | Approx 3 hours and 50 minutes |
| Wavelength | Primary wavelength : 450 nm Reference wavelength : 620 nm |
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 | 288 | 31.6 |
| 2 | 281 | 30.8 |
| 3 | 276 | 30.2 |
| 4 | 281 | 31.2 |
| 5 | 275 | 29.6 |
| mean | 280 | 30.7 |
| SD | 5.2 | 0.79 |
| CV(%) | 1.8 | 2.6 |
- [Result]
- The CV (%) of mouse serum was 1.8-2.6%, 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 | 200 | 32.0 | 5.22 |
| 2 | 193 | 32.2 | 5.16 |
| 3 | 192 | 33.0 | 4.91 |
| 4 | 195 | 32.7 | 4.89 |
| mean | 195 | 32.5 | 5.05 |
| SD | 3.6 | 0.46 | 0.17 |
| CV(%) | 1.8 | 1.4 | 3.4 |
- [Result]
- The CV (%) of mouse serum was 1.4-3.4%, indicating good reproducibility.
Dilution Linearity Test
Mouse serum and plasma (EDTA/heparin) were serially diluted twofold using the provided buffer and measured in duplicate to evaluate linearity.
Mouse serum
Mouse plasma (EDTA)
Mouse plasma (heparin)
- [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 and plasma (EDTA/heparin), 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 (%) |
|---|---|---|---|
| - | 8.70 | - | - |
| 20.0 | 27.9 | 19.2 | 96.0 |
| 80.0 | 95.5 | 86.8 | 109 |
| 165 | 174 | 165 | 100 |
| mean | 102 | ||
Mouse plasma (EDTA)
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 4.14 | - | - |
| 20.0 | 24.3 | 20.2 | 101 |
| 80.0 | 84.3 | 80.2 | 100 |
| 165 | 173 | 169 | 102 |
| mean | 101 | ||
Mouse plasma (heparin)
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 4.00 | - | - |
| 20.0 | 24.5 | 20.5 | 103 |
| 80.0 | 87.7 | 83.7 | 105 |
| 165 | 169 | 165 | 100 |
| mean | 103 | ||
- [Result]
- Good recovery rates were confirmed.
Example of Sample Measurement
Mouse serum and plasma (EDTA/heparin) samples were measured and compared using this kit and competitor Company A.
Mouse serum
| ID＼n | Fujifilm Wako(pg/mL) | Company A(pg/mL) |
|---|---|---|
| 11 | 30.1 | N.D. |
| 12 | 31.3 | N.D. |
| 13 | 5.73 | N.D. |
| 14 | 56.0 | N.D. |
| 15 | 19.3 | N.D. |
| mean | 28.5 | - |
Mouse plasma (EDTA)
| ID＼n | Fujifilm Wako(pg/mL) | Company A(pg/mL) |
|---|---|---|
| 11 | 1.85 | N.D. |
| 12 | 21.7 | N.D. |
| 13 | 38.0 | N.D. |
| 14 | 57.3 | N.D. |
| 15 | 5.85 | N.D. |
| mean | 30.7 | - |
Mouse plasma (heparin)
| ID＼n | Fujifilm Wako(pg/mL) | Company A(pg/mL) |
|---|---|---|
| 11 | 61.2 | N.D. |
| 12 | 18.3 | N.D. |
| 13 | 2.66 | 3.09 |
| 14 | 33.1 | N.D. |
| 15 | 77.6 | 4.46 |
| mean | 38.6 | - |
N.D.: Below the lower limit of quantification
Red: Values outside the quantification range; shown as reference
- [Result]
- This product was able to detect IFN-γ 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 using 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 mouse IFN-γ standard?
- The amount of purified water to be added varies by lot. Please reconstitute the mouse IFN-γ 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 IFN-γ 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
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Manufacturer Information
Alias
- 630-44701 (Previous code)
AKMIFNG-011 (Previous code)
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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