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LabAssay ™ Cholesterol

Manufacturer :
FUJIFILM Wako Shibayagi Corporation
Storage Condition :
Keep at 2-10 degrees C.
Molecular Weight :
0
GHS :
  • Structural Formula
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Product Number
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Price
Inventory
Distributor
635-50981
Manufacturer
LABCHO-M1
500Tests
List Price
214.00 USD

In stock in Japan

Document

Package Insert
Spectral Data
Certificate of Analysis
Calibration Certificate

Kit component

Kit component

Buffer 150 mL x 1 bottle
Chromogen Substrate for 150 mL x 1 bottle
Standard Solution 5 mL x 1 bottle

Overview

※For research purposes only with animal serum/plasma, Not for Diagnostic Use.

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 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)
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

04093456_img01.png

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 mouse and rat sample n=2

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

  • Rat serum, plasma
    04093456_img03.png
  • Mouse serum, plasma
    04093456_img04.png

Examples of measurement

  • 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
  • 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
  • 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
  • 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

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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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