LabAssay (TM) HDL-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 |
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Document
Kit component
For 100 tests
| Pretreatment | 20 mL/1 bottle |
|---|---|
| Reacting Solution | 8 mL/1 bottle |
| HDL-Cholesterol Standard | 2 bottles |
| Standard Diluent | 10 mL/1 bottle |
Product Overview
Cholesterol circulates in the body in particles called lipoproteins. There are different types of lipoprotein, among which HDL-cholesterol is responsible for transporting excess cholesterol to the liver, from tissues, eliminated cells, and blood, preventing increased blood cholesterol levels, and suppressing abnormal lipid metabolism and atherosclerosis. A decrease in HDL-cholesterol concentration is seen in coronary artery disease, hyperlipidemia and hypocholesterolemia, smoking, obesity, diabetes, and liver disease. HDL is an important protective factor against coronary heart disease (CHD), with hypo-HDL cholesterolemia one of the major risk factors for CHD.
LabAssay™ HDL-Cholesterol is a kit designed to measure HDL-cholesterol samples. With the use of a microplate, this kit provides a quick and convenient method for measuring HDL-cholesterol in samples. In addition, our kit do not require fractionation of lipoproteins by centrifugation.
Kit Performance
| Analysis sample | Human Serum/Plasma Mouse Serum/Plasma Rat Serum/Plasma |
|---|---|
| Calibration curve range | 6.25-200 mg/dL |
| Sample volume | 5 μL |
| Measurement duration | Approx. 20 min |
| Wavelength | Primary wavelength 600 nm Reference wavelength 700 nm |
Example of Calibration Curve
Assay Principle
1) The first reaction (elimination of non-HDL cholesterol)
The block polymer containing hydrophilic and hydrophobic moiety in Pretreatment selectively binds to HDL in the specimens, and protects it from enzyme (CHE, CO) reactions. CHE and CO react with non-HDL lipoprotein [chylomicron (CM), very low density lipoprotein (VLDL), low density lipoprotein (LDL)]. Hydrogen peroxide produced by the enzyme reactions with non-HDL cholesterol is decomposed to water by catalase in Pretreatment reagent.
2) The second reaction (color reaction of HDL-cholesterol)
When Reacting Solution is added, the cholesterol and its derivatives in HDL produce hydrogen peroxide by CHE and CO. Hydrogen peroxide produced by the enzyme reactions with HDL-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 HDL-cholesterol contained in the sample is determined by measuring the absorbance of the blue color.
Data
Repeatability (within run precision)
Repeatability was conducted by measuring five-fold assays of samples from human, mouse and rat at two concentrations with this product.
| n＼ID | Human sample | Mouse sample | Rat sample | |||
|---|---|---|---|---|---|---|
| ID1 (mg/dL) | ID2 (mg/dL) | ID1 (mg/dL) | ID2 (mg/dL) | ID1 (mg/dL) | ID2 (mg/dL) | |
| 1 | 82.3 | 33.3 | 72.0 | 21.2 | 55.0 | 21.4 |
| 2 | 82.5 | 32.2 | 73.2 | 20.7 | 53.5 | 21.5 |
| 3 | 82.2 | 32.9 | 72.1 | 21.2 | 54.0 | 21.1 |
| 4 | 81.9 | 33.4 | 71.4 | 22.1 | 54.3 | 22.8 |
| 5 | 82.2 | 32.9 | 72.8 | 21.3 | 55.8 | 22.1 |
| mean | 82.2 | 32.9 | 72.3 | 21.3 | 54.5 | 21.8 |
| SD | 0.217 | 0.472 | 0.707 | 0.505 | 0.898 | 0.676 |
| CV(%) | 0.3 | 1.4 | 1.0 | 2.4 | 1.6 | 3.1 |
[Result]
The CV (%) of human sample was 0.3-1.4%, mouse sample was 1.0-2.4%, and rat sample was 1.6-3.1%, indicating good repeatability.
Reproducibility (between run precision)
Reproducibility was conducted by measuring samples from human, mouse and rat at three concentrations each for four days with this product.
| Day＼ID | Human sample | Mouse sample | Rat sample | ||||||
|---|---|---|---|---|---|---|---|---|---|
| ID3 (mg/dL) | ID4 (mg/dL) | ID5 (mg/dL) | ID3 (mg/dL) | ID4 (mg/dL) | ID5 (mg/dL) | ID3 (mg/dL) | ID4 (mg/dL) | ID5 (mg/dL) | |
| 1 | 79.2 | 47.0 | 25.2 | 77.7 | 44.6 | 26.3 | 56.0 | 39.7 | 19.0 |
| 2 | 77.6 | 45.4 | 23.6 | 74.2 | 42.2 | 25.2 | 58.4 | 39.6 | 18.2 |
| 3 | 80.5 | 48.8 | 24.8 | 79.9 | 45.2 | 26.8 | 60.5 | 41.5 | 19.0 |
| 4 | 79.2 | 46.9 | 25.1 | 80.5 | 44.4 | 26.5 | 59.1 | 40.5 | 18.8 |
| mean | 79.1 | 47.0 | 24.7 | 78.1 | 44.1 | 26.2 | 58.5 | 40.3 | 18.8 |
| SD | 1.19 | 1.39 | 0.737 | 2.85 | 1.31 | 0.698 | 1.88 | 0.881 | 0.379 |
| CV(%) | 1.5 | 3.0 | 3.0 | 3.7 | 3.0 | 2.7 | 3.2 | 2.2 | 2.0 |
[Result]
The CV (%) of human sample was 1.5-3.0%, mouse sample was 2.7-3.7%, and rat sample was 2.0-3.2%, indicating good reproducibility.
Dilution Linearity Test
Serum and plasma (EDTA) from human, mouse and rat spiked with the standard, were serially diluted twofold with buffer in this kit and measured in duplicate to assess linearity.
[Result]
All measurement results showed good linearity within the measurement range
Spike and Recovery Test
Standard solutions at three concentrations were added to serum/plasma (EDTA) from human, mouse and rat. Next, spike recovery tests were conducted, with each measurement performed in duplicate.
Human sample
| Spiked amount (mg/dL) |
Measurement value (mg/dL) |
Recovery volume (mg/dL) |
Recovery rate (%) |
|
|---|---|---|---|---|
| Serum | - | 39.5 | - | - |
| 11.3 | 50.3 | 10.8 | 95.6 | |
| 22.5 | 61.8 | 22.3 | 99.1 | |
| 28.2 | 67.1 | 27.6 | 97.9 | |
| Average | 97.5 | |||
| Plasma (EDTA) |
- | 36.0 | - | - |
| 7.72 | 43.8 | 7.8 | 101 | |
| 15.4 | 50.4 | 14.4 | 93.5 | |
| 19.3 | 54.3 | 18.3 | 94.8 | |
| Average | 96.5 | |||
Mouse sample
| Spiked amount (mg/dL) |
Measurement value (mg/dL) |
Recovery volume (mg/dL) |
Recovery rate (%) |
|
|---|---|---|---|---|
| Serum | - | 11.7 | - | - |
| 11.8 | 22.5 | 10.8 | 91.5 | |
| 35.4 | 47.0 | 35.3 | 99.7 | |
| 59.1 | 67.4 | 55.7 | 94.2 | |
| Average | 95.2 | |||
| Plasma (EDTA) |
- | 10.5 | - | - |
| 9.34 | 19.2 | 8.7 | 93.1 | |
| 28.0 | 36.6 | 26.1 | 93.2 | |
| 46.7 | 54.9 | 44.4 | 95.1 | |
| Average | 93.8 | |||
Rat sample
| Spiked amount (mg/dL) |
Measurement value (mg/dL) |
Recovery volume (mg/dL) |
Recovery rate (%) |
|
|---|---|---|---|---|
| Serum | - | 15.6 | - | - |
| 15.6 | 30.0 | 14.4 | 92.3 | |
| 31.2 | 45.0 | 29.4 | 94.2 | |
| 39.1 | 51.4 | 35.8 | 91.6 | |
| Average | 92.7 | |||
| Plasma (EDTA) |
- | 13.0 | - | - |
| 13.2 | 25.3 | 12.3 | 93.2 | |
| 19.9 | 32.8 | 19.8 | 99.5 | |
| 33.1 | 44.0 | 31.0 | 93.7 | |
| Average | 95.4 | |||
[Result]
Good recovery rates were confirmed.
Measurement Using Actual Samples
Human sample
| ID＼n | Measurement value (mg/dL) |
mean | SD | CV (%) |
||
|---|---|---|---|---|---|---|
| 1 | 2 | |||||
| Serum | ID1 | 58.2 | 57.9 | 58.1 | 0.212 | 0.4 |
| ID2 | 44.1 | 45.2 | 44.7 | 0.778 | 1.7 | |
| ID3 | 45.7 | 45.7 | 45.7 | 0.00 | 0.0 | |
| ID4 | 35.3 | 34.3 | 34.8 | 0.707 | 2.0 | |
| ID5 | 34.9 | 34.3 | 34.6 | 0.424 | 1.2 | |
| Plasma (EDTA) |
ID1 | 104 | 105 | 104.5 | 0.707 | 0.7 |
| ID2 | 44.7 | 45.5 | 45.1 | 0.566 | 1.3 | |
| ID3 | 46.2 | 46.4 | 46.3 | 0.141 | 0.3 | |
| ID4 | 34.5 | 35.3 | 34.9 | 0.566 | 1.6 | |
| ID5 | 33.4 | 33.9 | 33.7 | 0.354 | 1.1 | |
Mouse sample (CD-1(ICR))
| ID＼n | Measurement value (mg/dL) |
mean | SD | CV (%) |
||
|---|---|---|---|---|---|---|
| 1 | 2 | |||||
| Serum | ID1 | 74.1 | 76.2 | 75.2 | 1.48 | 2.0 |
| ID2 | 71.9 | 71.7 | 71.8 | 0.141 | 0.2 | |
| ID3 | 77.2 | 80.3 | 78.8 | 2.19 | 2.8 | |
| ID4 | 86.5 | 87.7 | 87.1 | 0.849 | 1.0 | |
| ID5 | 68.2 | 69.3 | 68.8 | 0.778 | 1.1 | |
| Plasma (EDTA) |
ID1 | 40.6 | 40.3 | 40.5 | 0.212 | 0.5 |
| ID2 | 25.5 | 26.0 | 25.8 | 0.354 | 1.4 | |
| ID3 | 50.1 | 48.6 | 49.4 | 1.06 | 2.1 | |
| ID4 | 27.2 | 28.9 | 28.1 | 1.20 | 4.3 | |
| ID5 | 24.2 | 25.7 | 25.0 | 1.06 | 4.3 | |
Rat sample (S.D. rat)
| ID＼n | Measurement value (mg/dL) |
mean | SD | CV (%) |
||
|---|---|---|---|---|---|---|
| 1 | 2 | |||||
| Serum | ID1 | 22.5 | 20.5 | 21.5 | 1.41 | 6.6 |
| ID2 | 31.8 | 30.7 | 31.3 | 0.778 | 2.5 | |
| ID3 | 32.7 | 33.2 | 33.0 | 0.354 | 1.1 | |
| ID4 | 22.4 | 22.5 | 22.5 | 0.0707 | 0.3 | |
| ID5 | 25.5 | 25.8 | 25.7 | 0.212 | 0.8 | |
| Plasma (EDTA) |
ID1 | 13.2 | 13.3 | 13.3 | 0.0707 | 0.5 |
| ID2 | 13.3 | 13.1 | 13.2 | 0.141 | 1.1 | |
| ID3 | 13.1 | 11.4 | 12.3 | 1.20 | 9.8 | |
| ID4 | 13.6 | 13.6 | 13.6 | 0.00 | 0.0 | |
| ID5 | 15.4 | 16.4 | 15.9 | 0.707 | 4.4 | |
FAQ
About sample
- What types of samples should be used?
- Use serum or plasma samples that are as fresh as possible. Do not use samples that have undergone repeated freeze–thaw cycles, as lipoproteins may have become denatured.
- Do anticoagulants affect the assay?
- Heparin, citrate, and EDTA may all be used. When used at standard concentrations, they have little to no effect on the assay results.
- What should I do with samples that exceed the measurable range?
- For samples exceeding the upper limit of the assay range, dilute the sample with distilled water or an appropriate buffer prior to measurement. Multiply the result by the dilution factor to obtain the final value.
- How should serum samples with high triglyceride levels (1,000 mg/dL) be handled?
- For serum samples with triglyceride levels exceeding 1,000 mg/dL, dilute the sample with saline prior to measurement. Multiply the result by the dilution factor to obtain the final value.
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°C
- Plate mixer
- Microplate reader (with 600 nm/700 nm wavelength filter)
- What are the storage conditions and shelf-life guidelines for each reagent?
Pretreatment :
Ready to use. After opening the bottle, store at 2°C - 10°C and use within one month.Reacting Solution :
Ready to use. After opening the bottle, store at 2°C - 10°C and use within one month.Dilution series of standard solution :
The standard solution containing purified water should be stored at 2°C - 10°C and used within one month. Standard solutions prepared for each concentration should be used immediately and should not be stored.
- What is the amount of purified water to be added to the standard product?
- Find and check “Reconstitution of standard” on this product page. As the amount of purified water to be added varies by lot, be sure to check it for every lot.
Manufacturer Information
Alias
- LabAssay
For research use or further manufacturing use only. Not for use in diagnostic procedures.
Product content may differ from the actual image due to minor specification changes etc.
If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.
The prices are list prices in Japan.Please contact your local distributor for your retail price in your region.