LBIS™ Human MCP-1 (CCL2) ELISA Kit
- for Immunochemistry
- Manufacturer :
- FUJIFILM Wako Pure Chemical Corporation
- Storage Condition :
- Keep at 2-10 degrees C.
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96Tests
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Kit component
96 tests
| Antibody-coated Plate | 96 wells/1 plate |
|---|---|
| Human MCP-1 (CCL2) 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
MCP-1 (CCL2) is a member of the CC chemokine family and exhibits chemotactic activity toward monocytes and macrophages. Because MCP-1 and its receptor recruit these cells to sites of inflammation--thereby sustaining inflammatory responses--they are implicated in metabolic syndrome and cardiovascular diseases.
LBIS™ Human MCP-1 (CCL2) ELISA Kit is a sandwich ELISA kit designed for the quantitative measurement of human MCP-1 (CCL2). This kit enables highly sensitive quantification of MCP-1 (CCL2) in biological samples. The standard included in the kit is a recombinant protein produced in Escherichia coli.
Kit Performance
| Assay target | MCP-1 (CCL2) |
|---|---|
| Calibration curve range | 3.85 - 500 pg/mL |
| Analysis sample | Human Serum/Plasma (EDTA・Heparin)/Urine/ Culture medium (D-MEM・RPMI-1640 etc.)* |
| Sample amount | 20 μL (5x dilution) |
| Measurement duration | Approx 3 hours and 50 minutes |
| Wavelength | Primary wavelength : 450 nm Reference wavelength : 620 nm |
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 human serum samples at two concentrations.
| n＼ID | ID1 (pg/mL) | ID2 (pg/mL) |
|---|---|---|
| 1 | 284 | 49.8 |
| 2 | 280 | 49.9 |
| 3 | 279 | 50.2 |
| 4 | 281 | 49.4 |
| 5 | 279 | 48.7 |
| mean | 281 | 49.6 |
| SD | 2.1 | 0.58 |
| CV(%) | 0.74 | 1.2 |
- [Result]
- The CV (%) of human serum was 0.74-1.2%, 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 | 263 | 43.1 | 27.7 |
| 2 | 281 | 42.9 | 27.7 |
| 3 | 269 | 41.2 | 26.2 |
| 4 | 276 | 44.8 | 28.0 |
| mean | 272 | 43.0 | 27.4 |
| SD | 7.9 | 1.5 | 0.81 |
| CV(%) | 2.9 | 3.4 | 3.0 |
- [Result]
- The CV (%) of human serum was 2.9-3.4%, indicating good reproducibility.
Dilution Linearity Test
Human serum/plasma (EDTA/heparin) and urine samples were serially diluted twofold using the provided buffer and measured in duplicate to evaluate linearity.
Human serum
Human plasma (EDTA)
Human plasma (heparin)
Human urine
- [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), urine, and culture medium 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 (%) |
|---|---|---|---|
| - | 15.0 | - | - |
| 12.5 | 28.1 | 13.1 | 105 |
| 50 | 64.0 | 49.0 | 97.9 |
| 100 | 114 | 99.0 | 99.0 |
| 200 | 217 | 202 | 101 |
| mean | 101 | ||
Human plasma (EDTA)
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 15.5 | - | - |
| 12.5 | 28.3 | 12.8 | 102 |
| 50 | 64.5 | 49.0 | 98.0 |
| 100 | 108 | 92.5 | 92.5 |
| 200 | 204 | 189 | 94.5 |
| mean | 96.8 | ||
Human plasma (heparin)
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 13.9 | - | - |
| 12.5 | 25.9 | 12.0 | 96.0 |
| 50 | 63.3 | 49.4 | 98.8 |
| 100 | 109 | 95.1 | 95.1 |
| 200 | 206 | 192 | 96.0 |
| mean | 96.5 | ||
Human urine
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| - | 46.2 | - | - |
| 7.81 | 53.8 | 7.60 | 97.3 |
| 31.3 | 76.4 | 30.2 | 96.5 |
| 62.5 | 104 | 57.8 | 92.5 |
| 125 | 167 | 121 | 96.8 |
| mean | 95.8 | ||
Culture medium (SFM/CD/GIT/D-MEM/RPMI-1640)
| SFM | CD | GIT | D-MEM | RPMI-1640 | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Spiked amount (pg/mL) | Measurement value (pg/mL) | Recovery (%) | Spiked amount (pg/mL) | Measurement value (pg/mL) | Recovery (%) | Spiked amount (pg/mL) | Measurement value (pg/mL) | Recovery (%) | Spiked amount (pg/mL) | Measurement value (pg/mL) | Recovery (%) | Spiked amount (pg/mL) | Measurement value (pg/mL) | Recovery (%) |
| ― | 0 | ― | ― | 0 | ― | ― | 0 | ― | ― | 0 | ― | ― | 0 | ― |
| 12.5 | 12.6 | 101 | 12.5 | 13.5 | 108 | 12.5 | 12.0 | 95.8 | 12.5 | 12.0 | 95.7 | 12.5 | 13.5 | 108 |
| 50.0 | 49.8 | 99.7 | 50.0 | 52.0 | 104 | 50.0 | 50.5 | 101 | 50.0 | 49.7 | 99.5 | 50.0 | 52.5 | 105 |
| 100 | 99.5 | 99.5 | 100 | 97.6 | 97.6 | 100 | 103 | 103 | 100 | 94.8 | 94.8 | 100 | 97.3 | 97.3 |
| 200 | 200 | 100 | 200 | 196 | 97.9 | 200 | 197 | 98.3 | 200 | 188 | 94.0 | 200 | 197 | 98.3 |
| mean | 100 | mean | 102 | mean | 99.5 | mean | 96.0 | mean | 102 | |||||
- [Result]
- Good recovery rates were confirmed.
Correlation with conventional product
For Balb/c mouse serum/plasma (EDTA,heparin) samples, measurements were conducted using this kit and competitor product A. The measurement values obtained using both products were then compared.
Sample measurement value
Correlation
- [Result]
- Good correlation was confirmed.
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 human MCP-1 standard?
- The amount of purified water to be added varies by lot. Please reconstitute the human MCP-1 standard with the amount of purified water specified in the “Reconstitution of standard” section on this product page to prepare the original standard solution (5,000 pg/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 MCP-1 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
- 638-53411 (Previous code)
AKH-MCP1 (Previous code)
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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