Cholesterol assay kit

Manufactured by Applygen

Sourced in China

The Cholesterol assay kit is a laboratory tool used to measure the concentration of cholesterol in a sample. It provides a quantitative analysis of cholesterol levels, which is a key indicator of overall health and risk for certain medical conditions.

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Lab products found in correlation

Triglyceride assay kit, by Applygen (5 mentions) Mk886, by Cayman Chemical (1 mentions) Cholesterol assay kit, by Abcam (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Antibodies for western blotting, by Santa Cruz Biotechnology (1 mentions) Fetal bovine plasma, by Thermo Fisher Scientific (1 mentions) Avian myeloblastosis virus reverse transcriptase, by Promega (1 mentions) Cell culture medium, by Thermo Fisher Scientific (1 mentions) Oil red o, by Yeasen (1 mentions) T4 polynucleotide kinase, by Promega (1 mentions) Protein assay kit, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline tablets, by Takara Bio (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Oil red o, by Merck Group (1 mentions) Gpo pod, by Applygen (1 mentions)

12 protocols using cholesterol assay kit

Intracellular Cholesterol Quantification

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Intracellular total cholesterol (TC) and free cholesterol (FC) levels were detected by using cholesterol assay kits, an enzymatic assay technique, (Applygen Technologies, Beijing, China) according to the manufacturer's protocol.

Mei J., Xu F., Zhou Q., Zhang Y., Ji J, & Li M. (2022). Tetramethylpyrazine and Paeoniflorin Synergistically Attenuate Cholesterol Efflux in Macrophage Cells via Enhancing ABCA1 and ABCG1 Expression. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 4304790.

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Cholesterol Efflux Regulation by NAMPT

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RAW cells were grown to 60% confluence in 6-well plates, and then incubated with DMEM containing 1 μCi of [1, 2-³H] cholesterol per ml for 48 h. Percentages of cholesterol efflux to HDL were determined as mentioned above in RAW cells treatment with Ad-shNAMPT or Ad-GFP in the presence or absence of the selective inhibitor of PPARα (10 μM, MK886, Cayman Chemical Company). Total cellular cholesterol was measured using cholesterol assay kits purchased from Applygen Technologies Inc (Beijing, China). Expression of NAMPT, PPARα, LXRα, ABCA1 and ABCG1 were determined by Real-time RT-PCR and western blot analysis.

Li S., Wang C., Li K., Li L., Tian M., Xie J., Yang M., Jia Y., He J., Gao L., Boden G., Liu H, & Yang G. (2016). NAMPT knockdown attenuates atherosclerosis and promotes reverse cholesterol transport in ApoE KO mice with high-fat-induced insulin resistance. Scientific Reports, 6, 26746.

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Intrahepatic Lipid Quantification

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Intrahepatic triglycerides (TG) and cholesterol were measured by a triglyceride assay kit or cholesterol assay kit (Applygen Technologies Inc., Beijing, China). Samples and standards were then processed according to the manufacturer’s instructions. The final concentrations of triglycerides and cholesterol were corrected for protein content.

Zhou D., Pan Q., Shen F., Cao H.X., Ding W.J., Chen Y.W, & Fan J.G. (2017). Total fecal microbiota transplantation alleviates high-fat diet-induced steatohepatitis in mice via beneficial regulation of gut microbiota. Scientific Reports, 7, 1529.

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Isolation and Cholesterol Analysis of Cell Membranes

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To prepare cell membranes, previously reported methods were used with some modifications^{7 (link),56}. Briefly, CTLL2, CTLL2 treated with β-CD, CTLL2-PD1, and CTLL2-PD1 treated with β-CD were collected and disrupted in hypotonic lysing buffer (pH 7.4, 20 mM Tris-HCl, 10 mM KCl, 2 mM MgCl₂, Protease Inhibitor Cocktail) at 4 °C for 30 min. Destroyed cells in an ice bath using a probe ultrasonic instrument (150 W, 3 min). Then centrifuged at 20,000 × g for 20 min to remove organelles and large particles. The supernatant was centrifuged at 100,000 × g for 40 min to harvest cell membrane. The pellets were resuspended with buffer (10 mM Tris buffer, 1 mM EDTA (pH = 7.5)). The content of cholesterol in cell membrane was assayed using Cholesterol Assay Kit (ApplyGEN).

Li L., Zhang M., Li J., Liu T., Bao Q., Li X., Long J., Fu L., Zhang Z., Huang S., Liu Z, & Zhang L. (2023). Cholesterol removal improves performance of a model biomimetic system to co-deliver a photothermal agent and a STING agonist for cancer immunotherapy. Nature Communications, 14, 5111.

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Cellular Cholesterol Quantification

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Cells were cultured in DMEM (Gibco) with 10% lipid depleted FBS (S181L, Biowest) for 24 hours. The total cellular cholesterol content was measured using a commercial Cholesterol Assay Kit (E1005; Applygen Technologies, Inc.). Cholesterol staining was performed using a Cholesterol Assay Kit (ab133116, Abcam) according to the manufacturer's instructions.

Hu B., Zou T., Qin W., Shen X., Su Y., Li J., Chen Y., Zhang Z., Sun H., Zheng Y., Wang C.Q., Wang Z., Li T.E., Wang S., Zhu L., Wang X., Fu Y., Ren X., Dong Q, & Qin L.X. (2022). Inhibition of EGFR Overcomes Acquired Lenvatinib Resistance Driven by STAT3–ABCB1 Signaling in Hepatocellular Carcinoma. Cancer Research, 82(20), 3845-3857.

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In Vitro Evaluation of Injectable ART

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In this study, we used injectable ART (injection preparation, Guilin Pharma Corp, Guangxi, People’s Republic of China National Medicine Standard H10930195) with no endotoxin detected. We obtained 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and lipopolysaccharide (LPS) from Sigma-Aldrich (St. Louis, MO, USA); ox-LDL from Yiyuan (Guangzhou, Guangdong, China); and Oil Red O from Yeasen (Catalog No YB-002-1, Shanghai, China). Enhanced chemiluminescence (ECL) reagents were provided by Pierce, Inc. (Rockford, IL, USA). A cholesterol assay kit was provided by Applygen (Beijing, China). Human enzyme-linked immunosorbent assay (ELISA) kits for interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) were provided by Boster Ltd. (Wuhan, Hubei, China). Avian myeloblastosis virus (AMV) reverse transcriptase and T4 polynucleotide kinase were provided by Promega (Madison, WI, USA). Quantitative real-time polymerase chain reaction (qRT-PCR) master mix was provided by ToYoBo Ltd. (Osaka, Japan). All primers were synthesized by Invitrogen Ltd. (Shanghai, China). Cell culture medium and fetal bovine plasma were provided by Invitrogen (Carlsbad, CA, USA). The total protein extraction kit was provided by BestBio Ltd. (Shanghai, China). Antibodies for western blotting were provided by Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).

Qian Y., Xia L., Wei L, & Jiang W. (2021). Artesunate attenuates foam cell formation by enhancing cholesterol efflux. Annals of Translational Medicine, 9(17), 1379.

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Liver Lipid Profiling Protocol

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Liver levels of TGs and TC were detected by a triglyceride assay kit and a cholesterol assay kit, respectively (Applygen Technologies, Inc., Beijing, China), according to the manufacturer’s instructions. The final concentrations of TGs and TC were normalized to protein content.

Jiang D.X., Zhang J.B., Li M.T., Lin S.Z., Wang Y.Q., Chen Y.W, & Fan J.G. (2020). Prolyl endopeptidase gene disruption attenuates high fat diet-induced nonalcoholic fatty liver disease in mice by improving hepatic steatosis and inflammation. Annals of Translational Medicine, 8(5), 218.

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Intracellular Lipid Quantification

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Intracellular triglycerides and cholesterol were measured with the triglyceride assay kit (Applygen Technologies Inc.) and the cholesterol assay kit (Applygen Technologies Inc.), following the manufacturer’s protocols. The values were normalized to the total cellular protein content.

Yu P.W., Fu P.F., Zeng L., Qi Y.L., Li X.Q., Wang Q., Yang G.Y., Li H.W., Wang J., Chu B.B, & Wang M.D. (2022). EGCG Restricts PRRSV Proliferation by Disturbing Lipid Metabolism. Microbiology Spectrum, 10(2), e02276-21.

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Intracellular Lipid Quantification

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Lipid Metabolism Modulation Assay

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EPA and AA, dimethyl sulfoxide (DMSO), 3-(4, 5-dimethyl-thiazol-2-y)-2.5-diphenyl tetrazolium bromide (MTT), Phorbol-12-myristate-13-acetate(PMA) and Oil Red O were obtained from Sigma–Aldrich USA. Phosphate buffered saline (PBS) tablets were provided by Takara, Japan. Fetal bovine serum (FBS) were obtained from Invitrogen Corporation. Ox-LDL was purchased from YiYuan Biotech (Guangzhou, China). Fatty acids free BSA was obtained from Equitech-bio company (US). The ELISA kits of IL-6 and TNFα was purchased from Science Biotechnology Co. Ltd. (Yantai, China). The cholesterol assay kit was purchased from Applygen Technologies Inc. (Beijing, China). The protein assay kit was purchased from Thermo Fisher Scientific (Rockford, IL). Reagents and kits for RNA extraction and reverse transcription were obtained from Qiagen, USA.

Song Z., Xia H., Yang L., Wang S, & Sun G. (2018). Lowering the n-6/n-3 PUFAs ratio inhibits the formation of THP-1 macrophage-derived foam cell. Lipids in Health and Disease, 17, 125.

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