LabAssay (TM) Cholesterol
- Manufacturer :
- FUJIFILM Wako Shibayagi Corporation
- Storage Condition :
- Keep at 2-10 degrees C.
- Structural Formula
- Label
- Packing
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500Tests
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In stock in Japan |
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Document
Kit component
Kit component
| Buffer | 150 mL x 1 bottle |
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| Chromogen Substrate | for 150 mL x 1 bottle |
| Standard Solution | 5 mL x 1 bottle |
Overview
Introduction
Cholesterol concentration in serum is known to be closely related with production, absorption and dissimilation in liver and intestinal tract, and lipoprotein metabolism in blood.
LabAssay™ Cholesterol is able to measure total cholesterol in mouse and human serum by an enzyme reaction using N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium salt (DAOS).
Assay principle
Cholesterol esters in the sample are decomposed into free cholesterol and fatty acid by cholesterol esterase, when the sample reacts with chromogen reagent. The cholesterol is oxidized with existing free cholesterol by cholesterol oxidase, and simultaneously hydrogen peroxide is produced. The produced hydrogen peroxide let DAOS and 4-Aminoantipyrin oxidize and condensate quantitatively by peroxidase (HRP), which produces a blue pigment. Quantitation of total cholesterol in the sample can be made by measurement of the absorbance.
Performance
| Sample | Serum, plasma(Mouse, Rat, Human) |
|---|---|
| Standard curve range | 50 ~ 592.2 mg/dL |
| Measurement time | approx. 10 min. |
| Amount of sample | 2 μL |
| Measurement wavelength | 600 nm(main), 700 nm(sub) |
Standard Curve
Procedure
Perform the assay in the wells according to the following table scheme.
| Test | Standard | Blank | |
|---|---|---|---|
| Chromogen reagent | 300 μL | 300 μL | 300 μL |
| Sample | 2 μL | 2 μL | - |
| Mix well and incubate at 37 ℃ for 5 min. Measure the absorbance of the test sample and standard solution with the blank solution as the control. Main wavelength 600 nm / Sub wavelength 700 nm. | |||
Data
Spike-and-Recovery Test
Spike and Recovery Test of Cholesterol, a known amount of standard is spiked into in human, mouse, rat sample n=2
| Human Sample | Spike(mg/dL) | Mean(mg/dL) | Recovery(mg/dL) | Recovery(%) |
|---|---|---|---|---|
| Serum | 0.0 | 166.4 | ||
| 47.6 | 219.1 | 52.6 | 110.6% | |
| 99.7 | 276.2 | 109.7 | 110.0% | |
| 150.4 | 326.3 | 159.9 | 106.3% | |
| 201.3 | 378.3 | 211.9 | 105.2% | |
| Plasma (EDTA) | 0.0 | 130.2 | ||
| 47.6 | 181.0 | 50.8 | 106.6% | |
| 99.7 | 237.8 | 107.6 | 107.9% | |
| 150.4 | 280.5 | 150.3 | 100.0% | |
| 201.3 | 343.2 | 213.0 | 105.8% |
| Rat Sample | Spike(mg/dL) | Mean(mg/dL) | Recovery(mg/dL) | Recovery(%) |
|---|---|---|---|---|
| Serum | 0.0 | 69.9 | ||
| 47.6 | 121.5 | 51.6 | 108.3% | |
| 99.7 | 161.8 | 91.9 | 92.1% | |
| 150.4 | 220.8 | 150.8 | 100.3% | |
| 201.3 | 276.8 | 206.9 | 102.7% | |
| Plasma (EDTA) | 0.0 | 42.6 | ||
| 47.6 | 91.8 | 49.2 | 103.3% | |
| 99.7 | 137.6 | 95.0 | 95.3% | |
| 150.4 | 180.1 | 137.5 | 91.5% | |
| 201.3 | 235.8 | 193.2 | 96.0% |
| Mouse Sample | Spike(mg/dL) | Mean(mg/dL) | Recovery(mg/dL) | Recovery(%) |
|---|---|---|---|---|
| Serum | 0.0 | 86.1 | ||
| 47.6 | 136.1 | 50.0 | 105.0% | |
| 99.7 | 188.6 | 102.5 | 102.8% | |
| 150.4 | 239.3 | 153.2 | 101.9% | |
| 201.3 | 288.1 | 202.0 | 100.3% | |
| Plasma (EDTA) | 0.0 | 35.5 | ||
| 47.6 | 84.5 | 49.1 | 103.1% | |
| 99.7 | 134.4 | 98.9 | 99.2% | |
| 150.4 | 188.9 | 153.5 | 102.1% | |
| 201.3 | 230.6 | 195.1 | 96.9% |
Linearity-of-Dilution Test
- Human serum, plasma
- Rat serum, plasma
- Mouse serum, plasma
Examples of measurement
| Human serum, pooled, male | |||
|---|---|---|---|
| No. | Mean | SD | CV(%) |
| 1 | 0.687 | 0.001 | 0.2 |
| Human plasma K2EDTA, pooled, male | |||
| No. | Mean | SD | CV(%) |
| 1 | ※0.352 | 0.003 | 0.8 |
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20% Diluted Nomal Mouse serum, CD-1(ICR), male No. Mean (mEq/L) SD CV(%) 1 1.118 0.001 0.1 2 0.788 0.007 0.9 3 0.715 0.022 3.1 4 0.794 0.002 0.2 5 0.829 0.005 0.6 -
Nomal Mouse plasma K2EDTA, CD-1(ICR), male No. Mean SD CV(%) 1 ※0.309 0.002 0.6 2 ※0.320 0.002 0.6 3 ※0.205 0.004 2.0 4 ※0.179 0.008 4.6 5 ※0.305 0.017 5.6
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20% Diluted Nomal Rat serum, Sprague Dawley, male No. Mean SD CV(%) 1 ※0.300 0 0.0 2 ※0.371 0.004 1.1 3 ※0.289 0.017 5.8 4 ※0.365 0 0.1 5 ※0.382 0.029 7.6 -
Nomal Rat plasma K2EDTA, Sprague Dawley, male No. Mean SD CV(%) 1 ※0.333 0.004 1.2 2 ※0.356 0.003 0.8 3 ※0.325 0.020 6.0 4 ※0.218 0.005 2.6 5 0.515 0.010 1.9
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Nomal Canine plasma EDTA, Beagle, male No. Mean (mEq/L) SD CV(%) Age(years) 1 0.442 0.0033 0.7 4 2 0.427 0.0110 2.6 6 3 ※0.308 0.0037 1.2 10 4 0.478 0.0123 2.6 12 5 0.473 0.0050 1.0 6 6 ※0.367 0.0047 1.2 11 7 0.780 0.0133 1.7 10 8 0.411 0.0387 9.4 12 -
Nomal Canine serum, Beagle, male No. Mean (mEq/L) SD CV(%) Age(years) 1 0.656 0.0027 0.4 4 2 0.665 0.0320 4.8 6 3 0.425 0.0090 2.0 6 4 ※0.38 0.0057 1.5 11 5 0.882 0.0070 0.8 12 6 0.599 0.0060 1.1 6 7 1.006 0.0267 2.7 10 8 0.506 0.0060 1.2 4
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Nomal Feline plasma EDTA, Short hair, male No. Mean (mEq/L) SD CV(%) Age(years) 1 1.099 0.0103 0.9 0.5 2 ※0.174 0.0033 2.0 1 3 ※0.241 0.0043 1.7 3 4 ※0.320 0.0067 2.2 0.5 5 ※0.359 0.0060 1.7 3 6 ※0.197 0.0120 6.2 3 7 ※0.215 0.0240 11.1 0.5 8 ※0.131 0.0160 12.3 1 -
Nomal Feline serum, Short hair, male No. Mean (mEq/L) SD CV(%) Age(years) 1 0.717 0.0117 1.6 3 2 0.635 0.0100 1.6 3 3 1.004 0.0053 0.5 6 4 0.981 0.0093 0.9 4 5 0.636 0.0170 2.7 6 6 1.193 0.0267 2.3 6 7 1.022 0.0893 8.8 1 8 ※0.382 0.0157 4.1 1
References
- Kobayashi, Y. et al. : J. Pharmacogn. Nat. Prod., online (2015). Effects of Morus alba L. (Mulberry) Leaf Extract in Hypercholesterolemic Mice on Suppression of Cholesterol Synthesis *Extraction liquid of mouse Liver
- Gao, F. et al. : Evid. Based Complement. Alternat. Med., 2015, 801291 (2015). Effect of Keishibukuryogan on Genetic and Dietary Obesity Models *Extraction liquid of rat live
- Yoshioka, H. and Onosaka, S. : Fundam. Toxicol. Sci., 3, 151 (2016). Zinc sulfate pretreatment prevents carbon tetrachloride-induced lethal toxicity through metallothionein-mediated suppression of lipid peroxidation in mice *Mouse plasma
- Fujii, N. et al. : Aging Cell, 16, 508 (2017). Sterol regulatory element-binding protein-1c orchestrates metabolic remodeling of white adipose tissue by caloric restriction *Rat plasma
- Ushio, M. et al. : Am. J. Physiol. Endocrinol. Metab., 305, E293 (2013). Ezetimibe prevents hepatic steatosis induced by a high-fat but not a high-fructose diet *Extraction liquid of Mouse tissue
- Kobayashi, Y. et al. : Biosci. Biotechnol. Biochem., 74, 2385 (2010). Ameliorative Effects of Mulberry (Morus alba L.) Leaves on Hyperlipidemia in Rats Fed a High-Fat Diet: Induction of Fatty Acid Oxidation, Inhibition of Lipogenesis, and Suppression of Oxidative Stress *Extraction liquid of rat tissue
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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