LabAssay™ LDL-Cholesterol
- for Cellbiology
- Manufacturer :
- FUJIFILM Wako Pure Chemical Corporation
- Storage Condition :
- Keep at 2-10 degrees C.
- GHS :
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100Tests
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In stock in Japan |
※Check availability in the US with the distributor.
Document
Kit component
100 tests
| Pretreatment | 20 mL |
|---|---|
| Reacting Solution | 8 mL |
| LDL-Cholesterol Standard | 2 vials |
| Standard Diluent | 10 mL |
Product Overview
The cholesterol carried by Low Density Lipoprotein (LDL) termed LDL-cholesterol. LDL carries cholesterol produced in the liver throughout the body, and when present in large amounts in the blood, it deposits and accumulates in blood vessel walls, causing atherosclerosis and resulting in myocardial infarction and cerebral infarction.
In humans, the normal range of LDL-cholesterol is less than 140 mg/dL, and more than 140 mg/dL is considered to reflect hyper-LDL cholesterolemia.
LabAssay™ LDL-Cholesterol is a kit for the determination of LDL-cholesterol in blood (serum and plasma). With the use of a microplate, this kit provides a quick and convenient method for measuring LDL-cholesterol in samples. In addition, our kit do not require fractionation of lipoproteins by centrifugation.
Kit Performance
| Sample | Serum, Plasma |
|---|---|
| Target animals | Mouse, Rat |
| Calibration curve range | 9.38-300 mg/dL |
| Sample volume | 5 μL |
| Measurement duration | Approx. 20 min |
| Wavelength | Primary wavelength 600 nm Reference wavelength 700 nm |
Example of Calibration Curve
Principle
1) The first reaction（elimination of non-LDL cholesterol）
The block polymer containing hydrophilic and hydrophobic moiety in Pretreatment selectively binds to LDL in the specimens, and protects it from enzyme (CHE, CO) reactions. CHE and CO react with non-LDL lipoprotein [chylomicron (CM), very low density lipoprotein (VLDL), high density lipoprotein (HDL)]. Hydrogen peroxide produced by the enzyme reactions with non-LDL cholesterol is decomposed to water by catalase in Pretreatment reagent.
2) The second reaction（color reaction of LDL-cholesterol)
When Reacting Solution is added, the cholesterol and its derivatives in LDL produce hydrogen peroxide by CHE and CO. Hydrogen peroxide produced by the enzyme reactions with LDL-cholesterol yields a blue color pigment upon oxidative condensation with N-(3-sulfopropyl)-3-methoxy-5-methylaniline (HMMPS) and 4-aminoantipyrine in the presence of peroxidase (POD). The amount of LDL-cholesterol contained in the sample is determined by measuring the absorbance of the blue color.
Data
Precision of assay (within assay variation)
Reproducibility was confirmed by performing quintuplicate assays of serum samples of two concentrations each from humans, mice, and rats with this product.
| n＼ID | Human | Mouse | Rat | |||
|---|---|---|---|---|---|---|
| ID1 (mg/dL) | ID2 (mg/dL) | ID1 (mg/dL) | ID2 (mg/dL) | ID1 (mg/dL) | ID2 (mg/dL) | |
| 1 | 160 | 46.0 | 141 | 48.3 | 119 | 33.3 |
| 2 | 156 | 43.8 | 138 | 47.9 | 114 | 34.1 |
| 3 | 158 | 44.7 | 138 | 49.5 | 119 | 33.3 |
| 4 | 166 | 44.5 | 139 | 48.5 | 112 | 34.9 |
| 5 | 159 | 45.7 | 136 | 50.6 | 113 | 33.3 |
| mean | 160 | 44.9 | 138 | 49.0 | 115 | 33.8 |
| SD | 3.8 | 0.90 | 1.8 | 1.1 | 3.4 | 0.72 |
| CV(%) | 2.4 | 2.0 | 1.3 | 2.2 | 2.9 | 2.1 |
[Result]
The CV (%) of human serum was 2.0-2.4%, mouse serum was 1.3-2.2%, and rat serum was 2.1-2.9%, indicating good reproducibility.
Reproducibility (between assay variation)
Triplicate measurements were performed on human, mouse, and rat serum samples at three concentrations each for four days with this product.
| Day＼ID | Human | Mouse | Rat | ||||||
|---|---|---|---|---|---|---|---|---|---|
| ID1 (mg/dL) | ID2 (mg/dL) | ID3 (mg/dL) | ID1 (mg/dL) | ID2 (mg/dL) | ID3 (mg/dL) | ID1 (mg/dL) | ID2 (mg/dL) | ID3 (mg/dL) | |
| 1 | 125 | 88.1 | 41.8 | 119 | 91.4 | 48.4 | 130 | 78.7 | 45.2 |
| 2 | 124 | 87.0 | 41.1 | 119 | 88.1 | 45.0 | 129 | 76.1 | 42.9 |
| 3 | 130 | 91.5 | 41.7 | 118 | 88.0 | 48.2 | 126 | 74.2 | 49.7 |
| 4 | 125 | 89.5 | 39.8 | 126 | 95.1 | 50.4 | 131 | 74.5 | 49.4 |
| mean | 126 | 89.0 | 41.1 | 121 | 90.7 | 48.0 | 129.0 | 75.9 | 46.8 |
| SD | 2.7 | 1.9 | 0.92 | 3.7 | 3.4 | 2.2 | 2.2 | 2.1 | 3.3 |
| CV(%) | 2.1 | 2.2 | 2.2 | 3.1 | 3.7 | 4.7 | 1.7 | 2.7 | 7.1 |
[Result]
The CV (%) of human serum was 2.1-2.2%, mouse serum was 3.1-4.7%, and rat serum was 1.7-7.1%, indicating good reproducibility.
Dilution linearity test
Using the standard diluent supplied with the product, human serum/plasma (EDTA), mouse serum/plasma (EDTA), and rat serum/plasma (EDTA) at two different concentrations each, were subjected to a 2-fold serial dilution with sample buffer and measured to confirm linearity (measured in duplicates).
[Result]
Good linearity was confirmed for all samples within the calibration curve range.
Spiked recovery test
Standard solutions at three concentrations were added to human serum/EDTA plasma, mouse serum/EDTA plasma and rat serum/EDTA plasma. Next, spike-recovery tests were conducted, with each measurement performed in duplicate.
Human
| Amount spiked (mg/dL) | Measured value (mg/dL) | Recovery volume (mg/dL) | Recovery rate (%) | |
|---|---|---|---|---|
| Serum | - | 75.2 | - | - |
| 7.12 | 82.3 | 7.10 | 100 | |
| 28.5 | 103 | 27.8 | 97.5 | |
| 35.6 | 112 | 36.8 | 103 | |
| Plasma (EDTA) |
- | 84.5 | - | - |
| 10.6 | 93.8 | 9.30 | 87.7 | |
| 21.2 | 107 | 22.5 | 106 | |
| 26.5 | 112 | 27.5 | 104 |
Mouse
| Amount spiked (mg/dL) | Measured value (mg/dL) | Recovery volume (mg/dL) | Recovery rate (%) | |
|---|---|---|---|---|
| Serum | - | 86.6 | - | - |
| 15.9 | 102 | 15.4 | 96.9 | |
| 23.8 | 112 | 25.4 | 107 | |
| 39.7 | 126 | 39.4 | 99.2 | |
| Plasma (EDTA) |
- | 97.7 | - | - |
| 14.4 | 112 | 14.3 | 99.3 | |
| 28.9 | 126 | 28.3 | 97.9 | |
| 36.1 | 133 | 35.3 | 97.8 |
Rat
| Amount spiked (mg/dL) | Measured value (mg/dL) | Recovery volume (mg/dL) | Recovery rate (%) | |
|---|---|---|---|---|
| Serum | - | 14.6 | - | - |
| 13.7 | 28.7 | 14.1 | 103 | |
| 41.0 | 53.0 | 38.4 | 93.7 | |
| 68.4 | 77.1 | 62.5 | 91.4 | |
| Plasma (EDTA) |
- | 16.2 | - | - |
| 16.0 | 31.9 | 15.7 | 98.1 | |
| 48.1 | 64.4 | 48.2 | 100 | |
| 80.2 | 90.6 | 74.4 | 92.8 |
[Result]
Good recovery rates was confirmed for all samples.
Measurement examples
Human serum/plasma, mouse (ICR) serum/plasma samples, and rat (SD) serum/plasma were measured with this product.
Human
| No. | Measured value (mg/dL) | mean (mg/dL) | SD | CV(%) | ||
|---|---|---|---|---|---|---|
| Serum | 1 | 189 | 187 | 188 | 1.4 | 0.75 |
| 2 | 88.8 | 93.2 | 91 | 3.1 | 3.4 | |
| 3 | 85.7 | 85.7 | 85.7 | 0 | 0 | |
| 4 | 99 | 93.9 | 96.5 | 3.6 | 3.7 | |
| 5 | 95.8 | 94 | 94.9 | 1.3 | 1.3 | |
| Plasma (EDTA) |
1 | 122 | 121 | 122 | 0.71 | 0.58 |
| 2 | 88.3 | 88 | 88.2 | 0.21 | 0.24 | |
| 3 | 85.9 | 84.4 | 85.2 | 1.1 | 1.2 | |
| 4 | 93.9 | 97.2 | 95.6 | 2.3 | 2.4 | |
| 5 | 95.1 | 95.4 | 95.3 | 0.21 | 0.22 | |
Mouse
| No. | Measured value (mg/dL) | mean (mg/dL) | SD | CV(%) | ||
|---|---|---|---|---|---|---|
| Serum | 1 | 32.6 | 33.4 | 33 | 0.57 | 1.7 |
| 2 | 33.6 | 31.3 | 32.5 | 1.6 | 5 | |
| 3 | 33.4 | 32.4 | 32.9 | 0.71 | 2.1 | |
| 4 | 34.5 | 33.4 | 34 | 0.78 | 2.3 | |
| 5 | 36.1 | 34.1 | 35.1 | 1.4 | 4 | |
| Plasma (EDTA) |
1 | 21.2 | 20.8 | 21 | 0.28 | 1.3 |
| 2 | 16 | 17.7 | 16.9 | 1.2 | 7.1 | |
| 3 | 25.5 | 21.2 | 23.4 | 3 | 13 | |
| 4 | 19.1 | 20.3 | 19.7 | 0.85 | 4.3 | |
| 5 | 18.9 | 19.6 | 19.3 | 0.49 | 2.6 | |
Rat
| No. | Measured value (mg/dL) | mean (mg/dL) | SD | CV(%) | ||
|---|---|---|---|---|---|---|
| Serum | 1 | 43.3 | 37.9 | 40.6 | 3.8 | 9.4 |
| 2 | 44.4 | 42.3 | 43.4 | 1.5 | 3.4 | |
| 3 | 42.7 | 44 | 43.4 | 0.92 | 2.1 | |
| 4 | 44.9 | 43.2 | 44.1 | 1.2 | 2.7 | |
| 5 | 50 | 51.1 | 50.6 | 0.78 | 1.5 | |
| Plasma (EDTA) |
1 | 30.7 | 30.1 | 30.4 | 0.42 | 1.4 |
| 2 | 33.7 | 32.8 | 33.3 | 0.64 | 1.9 | |
| 3 | 39.2 | 36.5 | 37.9 | 1.9 | 5 | |
| 4 | 30.7 | 30.9 | 30.8 | 0.14 | 0.46 | |
| 5 | 33.9 | 36.7 | 35.3 | 2 | 5.6 | |
FAQ
About sample
- What specimens should I use?
- Use fresh specimens. Do not use specimens after repeated freeze-thawing as this may denature lipoproteins.
- Which anticoagulants can I use?
- Anticoagulants such as heparin, citrate and EDTA do not significantly influence the assay when used in normal amounts.
- What should I do with samples that exceed the measurable range?
- Dilute specimen with saline and repeat the assay if the measured value exceeds the measurable range, and multiply the result by the dilution factor.
- How should I do when triglyceride in the specimen exceeds 1,000 mg/dL?
- When triglyceride in the specimen exceeds 1,000 mg/dL, dilute the specimen with saline and multiply the result by the dilution factor.
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.
- 96-well microplate (transparent type)
- Micropipette
- Incubator maintained at 37℃
- Plate mixer
- Microplate reader (with 600 nm/700 nm wavelength filter)
- What is the amount of purified water to be added to the standard product?
- Because the volume of distilled water to be added to the standard differs depending on the lot, see the specified volume in a separate sheet.
Overview / Applications
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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