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Total Cholesterol (TC) Colorimetric Assay Kit (Single Reagent, COD-PAP Method)

Cat.No.:E-BC-K109-M
Detection instrument: Microplate reader(495-525 nm)

Detection method: Colorimetric method

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Detection principle

Total cholesterol includes free cholesterol and cholesterol esters. Cholesterol ester can be hydrolyzed by cholesterol esterase to produce cholesterol and free fatty acid. Cholesterol is oxidized by cholesterol oxidase to produce △4-cholestenone and hydrogen peroxide. In the presence of 4-aminoamylpyridine and phenol, hydrogen peroxide catalyze peroxidase to form red quinone compounds of benzoquinone imine phenizone.The color depth of the generated quinone is directly proportional to the cholesterol content. The absorbance values of the standard tube and the sample tube are measured respectively, and the cholesterol content in the sample can be calculated.

Performance characteristics

Synonyms	TC
Sample type	Serum,plasma,tissue
Sensitivity	0.29 mmol/L
Detection range	0.29-25.85 mmol/L
Detection method	Colorimetric method
Assay type	Quantitative
Assay time	20 min
Precision	Average inter-assay CV: 8.300%Average intra-assay CV: 3.100%
Other instruments required	Micropipettor, Incubator, Centrifuge
Other reagents required	Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4), Anhydrous ethanol
Storage	2-8℃
Valid period	12 months

Images

The concentration of cholesterol in THP-1 cell was significantly increased in ox-LDL group. (***P<0.001)

C Zhu et al found that TRIM64 can promote pyroptosis, foam cell formation and inflammation by activating NF-κB. Total cholesterol (TC) levels in human THP-1 cell was determined using TC colorimetric assay kit (E-BC-K109-M).

Dilution of sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.29-25.85 mmol/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type	Dilution factor
Human serum	1
Mouse serum	1
Rat plasma	1
10% Mouse liver tissue homogenate	1
10% Mouse kidney tissue homogenate	1
10% Mouse heart tissue homogenate	1

Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4) for serum (plasma) samples; The diluent is anhydrous ethanol for tissue samples.

Troubleshooting

Collection and treatment of common samples for metabolism assay detection

Questions and Answers of Metabolism Assay Kits

Citations

Advanced Science (2023) IF: 17.521

Phosphomevalonate Kinase Controls β-Catenin Signaling via the Metabolite 5-Diphosphomevalonate

DOI: 10.1002/advs.202204909
PMID: 36808719	Sample: Serum

CELL PROLIFERATION (2022) IF: 8.7550

Hypothalamic Hnscr regulates glucose balance by mediating central inflammation and insulin signal

DOI: 10.1111/cpr.13332
	Sample: liver

Frontiers in Endocrinology (2022) IF: 6.055

Total Flavonoids of Drynariae Rhizoma Improve Glucocorticoid-Induced Osteoporosis of Rats: UHPLC-MS-Based Qualitative Analysis, Network Pharmacology Strategy and Pharmacodynamic Validation

DOI: 10.3389/fendo.2022.920931
	Sample: Serum

Food & Function (2023) IF: 6.317

Analysis of the mechanism of action of quercetin in the treatment of hyperlipidemia based on metabolomics and intestinal flora

DOI: 10.1039/D2FO03509J
	Sample: serum

EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY (2021) IF: 6.514

Design, synthesis, and biological evaluation of a novel dual peroxisome proliferator-activated receptor alpha/delta agonist for the treatment of diabetic kidney disease through anti-inflammatory mechanisms

DOI: 10.1016/j.ejmech.2021.113388
PMID: 33784603	Sample: serum

Frontiers in Cell and Developmental Biology (2021) IF: 6.684

N6-Methyladenosine Methyltransferase METTL3 Promotes Angiogenesis and Atherosclerosis by Upregulating the JAK2/STAT3 Pathway via m6A Reader IGF2BP1.

DOI: 10.3389/fcell.2021.731810
	Sample: plasma

Nutrients (2022) IF: 6.706

Ascophyllum nodosum and Fucus vesiculosus Extracts Improved Lipid Metabolism and Inflammation in High-Energy Diet–Induced Hyperlipidemia Rats

DOI: 10.3390/nu14214665
	Sample: liver

CELL BIOLOGY AND TOXICOLOGY (2022) IF: 6.819

TRIM64 promotes ox-LDL-induced foam cell formation, pyroptosis, and inflammation in THP-1-derived macrophages by activating a feedback loop with NF-κB via IκBα ubiquitination

DOI: 10.1007/s10565-022-09768-4
PMID: 36229750	Sample: THP-1 cells

JOURNAL OF ETHNOPHARMACOLOGY (2022) IF: 5.195

Integrating metabolomics and network pharmacology to investigate Panax japonicus prevents kidney injury in HFD/STZ-induced diabetic mice

DOI: 10.1016/j.jep.2022.115893
	Sample: serum

COLLOIDS AND SURFACES B-BIOINTERFACES (2021) IF: 5.268

AFM detects the effects of acidic condition on the size and biomechanical properties of native/oxidized low-density lipoprotein

DOI: 10.1016/j.colsurfb.2021.112053
PMID: 34438294	Sample: plasma

MOLECULAR NUTRITION & FOOD RESEARCH (2020) IF: 5.309

Gymnaster Koraiensis Extract Alleviated Metabolic Syndrome Symptoms and Stimulated UCP1‐Independent Energy Consumption via AMPK Activation in White Adipose Tissue

DOI: 10.1002/mnfr.202000490
	Sample: Serum

Food Bioscience (2022) IF: 5.318

Ameliorative effects of monascin from red mold rice on alcoholic liver injury and intestinal microbiota dysbiosis in mice

DOI: 10.1016/j.fbio.2022.102079
	Sample: serum,liver

Frontiers in Endocrinology (2021) IF: 5.555

Metabolic Consequences of Neuronal HIF1α-Deficiency in Mediobasal Hypothalamus in Mice

DOI: 10.3389/fendo.2021.668193
PMID: 34733235	Sample: serum

Foods (2022) IF: 5.561

Monascuspiloin from Monascus-Fermented Red Mold Rice Alleviates Alcoholic Liver Injury and Modulates Intestinal Microbiota

DOI: 10.3390/foods11193048
PMID: 36230124	Sample: liver,fecal,serum

AMERICAN JOURNAL OF PATHOLOGY (2022) IF: 5.77

Paraoxonase 1 Ameliorates Renal Lipotoxicity by Activating Lipophagy and Inhibiting Pyroptosis

DOI: 10.1016/j.ajpath.2022.07.011
PMID: 35963464	Sample: plasma

ACS Omega (2022) IF: 4.132

Beneficial Effects of lncRNA-UC.360+ shRNA on Diabetic Cardiac Sympathetic Damage via NLRP3 Inflammasome-Induced Pyroptosis in Stellate Ganglion

DOI: 10.1021/acsomega.2c03619
PMID: 35967043	Sample: serum

JOURNAL OF ETHNOPHARMACOLOGY (2022) IF: 4.36

Zexie Tang targeting FKBP38/mTOR/SREBPs pathway improves hyperlipidemia

DOI: 10.1016/j.jep.2022.115101
PMID: 35151834	Sample: serum

TOXICOLOGY LETTERS (2021) IF: 4.374

Alpha-lipoic acid attenuates silica-induced pulmonary fibrosis by improving mitochondrial function via AMPK/PGC1α pathway activation in C57BL/6J mice

DOI: 10.1016/j.toxlet.2021.07.003
PMID: 34252510	Sample: serum

Journal of Functional Foods (2021) IF: 4.451

Heracleum dissectum Ledeb. ethanol extract attenuates metabolic syndrome symptoms in high-fat diet-induced obese mice by activating adiponectin/AMPK signaling

DOI: 10.1016/j.jff.2021.104581
	Sample: blood

JOURNAL OF CELLULAR AND MOLECULAR MEDICINE (2020) IF: 4.486

Isoliquiritigenin ameliorates caerulein‐induced chronic pancreatitis by inhibiting the activation of PSCs and pancreatic infiltration of macrophages

DOI: 10.1111/jcmm.15498
	Sample: Serum

JOURNAL OF PSYCHIATRIC RESEARCH (2022) IF: 4.791

Comparison of cognitive impairments with lipid profiles and inflammatory biomarkers in unipolar and bipolar depression

DOI: 10.1016/j.jpsychires.2022.04.002
PMID: 35429740	Sample: serum

Diabetes Metabolic Syndrome and Obesity-Targets and Therapy (2022) IF: 3.168

Serum Levels of Free Fatty Acids in Obese Mice and Their Associations with Routine Lipid Profiles

DOI: 10.2147/DMSO.S348800
PMID: 35140491	Sample: serum

ANIMAL FEED SCIENCE AND TECHNOLOGY (2021) IF: 3.247

Coating beef tallow with calcium soap improves dietary pellet quality and promotes an effective diet-induced obese mouse model

DOI: 10.1016/j.anifeedsci.2021.115008
	Sample: plasma

MEAT SCIENCE (2019) IF: 3.483

Effects of naturally-produced lovastatin on carcass characteristics, muscle physico-chemical properties and lipid oxidation and cholesterol content in goats

DOI: 10.1016/j.meatsci.2019.04.008
	Sample: serum

AQUACULTURE RESEARCH (2021) IF: 2.082

Quillaja saponaria and/or Yucca schidigera ameliorate water quality, growth performance, blood health, intestine and gills histomorphology of Nile tilapia, Oreochromis niloticus

DOI: 10.1111/are.15474
	Sample: serum

THERIOGENOLOGY (2020) IF: 2.094

Diet-induced obesity is associated with altered expression of sperm motility-related genes and testicular post-translational modifications in a mouse model

DOI: 10.1016/j.theriogenology.2020.09.023
PMID: 32980686	Sample: Serum

Journal of Natural Medicines (2021) IF: 2.343

Dehydroabietic acid improves nonalcoholic fatty liver disease through activating the Keap1/Nrf2-ARE signaling pathway to reduce ferroptosis

DOI: 10.1007/s11418-021-01491-4
PMID: 33590347	Sample: Tissue homogenate(liver)

Journal of Developmental Origins of Health and Disease (2020) IF: 2.456

Neonatal zingerone protects against the development of high-fructose diet-induced metabolic syndrome in adult Sprague-Dawley rats

DOI: 10.1017/S2040174420000525
	Sample: plasma

NUTRITION (2016) IF: 2.839

Dietary sweet cherry anthocyanins attenuates diet-induced hepatic steatosis by improving hepatic lipid metabolism in mice

DOI: 10.1016/j.nut.2016.01.007
	Sample: liver tissue

International Journal of Clinical and Experimental Pathology (2018) IF: 1.396

Pitavastatin attenuates atherosclerosis by suppressing NF-κB signaling in a high-cholesterol diet plus balloon catheter injury rabbit model


PMID: 31938118	Sample: serum

TOXICOLOGY AND INDUSTRIAL HEALTH (2019) IF: 1.635

Toxic effects of subchronic oral acetamiprid exposure in rats:

DOI: 10.1177/0748233719893203
	Sample: plasma

TOXICOLOGY AND INDUSTRIAL HEALTH (2020) IF: 1.708

Elevated, sustained, and yet reversible biotoxicity effects of lead on cessation of exposure: Melatonin is a potent therapeutic option:

DOI: 10.1177/0748233720937199
	Sample: Plasma

BMC Veterinary Research (2020) IF: 1.835

Tea polyphenols attenuate liver inflammation by modulating obesity-related genes and down-regulating COX-2 and iNOS expression in high fat-fed dogs

DOI: 10.1186/s12917-020-02448-7
PMID: 32641048	Sample: Serum,Tissue homogenate

Veterinary World (2020)

Cinnamomum burmannii (Nees & T. Nees) Blume and Eleutherine palmifolia (L.) Merr. extract combination ameliorate lipid profile and heart oxidative stress in hyperlipidemic mice

DOI: 10.14202/vetworld.2020.1404-1409
PMID: 32848317	Sample: Serum

Journal of Advanced Pharmaceutical Technology & Research (2020)

Antihyperlipidemic effects of apple peel extract in high-fat diet-induced hyperlipidemic rats

DOI: 10.4103/japtr.JAPTR_28_20
	Sample: serum

Revista Brasileira de Medicina do Esporte (2022) IF: 0.652

IMPACTS OF AEROBIC EXERCISE ON THE OBESITY OF ADOLESCENTS AND THEIR LIPID METABOLISM

DOI: 10.1590/1517-8692202329012022_0163
	Sample: serum

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Reviews/Q&A

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Reviews
Q&A

M*****aSubmitted [ Oct 10 2022 ]

Asked: The homogenate medium of TC described in the protocol is isopropanol. Can it be changed to normal saline?`

adminSubmitted [ Oct 10 2022 ]

Answered: I really don't recommend that, the effect of normal saline homogenate is not good, leading to undetectable results.

M*****aSubmitted [ Sep 27 2022 ]