LBIS™ Human VEGF 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 VEGF 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
Vascular Endothelial Growth Factor (VEGF) is a cytokine that acts specifically on vascular endothelial cells. It promotes angiogenesis and enhances vascular permeability. VEGF has been implicated in diseases such as cancer, rheumatoid arthritis, arteriosclerosis, and retinopathy.
LBIS™ Human VEGF ELISA Kit is a sandwich ELISA kit for the quantitative measurement of human VEGF. This kit enables highly sensitive quantification of VEGF. The standard included in the kit is a recombinant protein produced in Escherichia coli.
Kit Performance
| Assay target | VEGF (VEGF-A165) |
|---|---|
| Calibration curve range | 1.10 - 800 pg/mL |
| Analysis sample | Human Serum/Plasma (EDTA, heparin) /Cell culture supernatant* |
| Sample amount | 100 μL |
| Measurement duration | Approx 4 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 human serum samples at two concentrations.
| n＼ID | ID1 (pg/mL) | ID2 (pg/mL) |
|---|---|---|
| 1 | 29.7 | 37.8 |
| 2 | 28.3 | 36.5 |
| 3 | 27.7 | 35.0 |
| 4 | 29.3 | 35.4 |
| 5 | 28.8 | 35.4 |
| mean | 28.8 | 36.0 |
| SD | 0.79 | 1.1 |
| CV(%) | 2.8 | 3.2 |
- [Result]
- The CV (%) of human serum was 2.8-3.2%, indicating good repeatability.
Reproducibility (between run precision)
Reproducibility was conducted by measuring human serum samples at three concentrations in triplicate each for four days.
| Day＼ID | ID3 (pg/mL) | ID4 (pg/mL) | ID5 (pg/mL) |
|---|---|---|---|
| 1 | 279 | 86.1 | 11.9 |
| 2 | 271 | 84.5 | 12.5 |
| 3 | 268 | 88.5 | 11.2 |
| 4 | 269 | 88.1 | 11.5 |
| mean | 272 | 86.8 | 11.8 |
| SD | 5.0 | 1.9 | 0.56 |
| CV(%) | 1.8 | 2.1 | 4.8 |
- [結果]
- The CV (%) of human serum was 1.8-4.8%, indicating good reproducibility.
Dilution Linearity Test
Human serum and plasma, 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
Cell culture supernatant (COLO201)
- [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, 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 (%) |
|---|---|---|---|
| 0 | 50.6 | - | - |
| 89.0 | 145 | 94.4 | 106 |
| 189 | 237 | 186 | 98.4 |
| 388 | 456 | 405 | 104 |
| mean | 103 | ||
Human plasma
| Spiked amount (pg/mL) |
Measurement value (pg/mL) |
Recovery amount (pg/mL) |
Recovery (%) |
|---|---|---|---|
| 0 | 42.1 | - | - |
| 97.5 | 134 | 91.9 | 94.3 |
| 197 | 240 | 198 | 100 |
| 396 | 420 | 378 | 95.5 |
| mean | 96.6 | ||
- [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
Human plasma (EDTA)
Human plasma (heparin)
Bars are omitted when the measured value falls outside the range of quantification.
- [Result]
- For most human serum samples, this product delivered results comparable to those obtained with the other manufacturer's product. In both serum and plasma, VEGF was also measurable with this product in samples in which the other manufacturer's product did not detect VEGF.
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°C 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 VEGF standard?
- The amount of purified water to be added varies by lot. Please reconstitute the human VEGF standard with the amount of purified water specified in the “Reconstitution of standard” section on this product page to prepare the original standard solution (8 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°C 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°C.
Human VEGF 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°C. 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
- 631-40831 (Previous code)
AKH-VEGF (Previous code)
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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