LabAssay (TM) Cholesterol

※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

Standard Curve

Procedure

Perform the assay in the wells according to the following table scheme.

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

Linearity-of-Dilution Test

• Rat serum, plasma
  
  
• Mouse serum, plasma
  
  

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

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