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Mak043

Manufactured by Merck Group
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Sourced in United States

The MAK043 is a laboratory instrument designed for scientific research and analysis. It is a precision piece of equipment used to perform specific tasks in a controlled environment. The core function of the MAK043 is to provide accurate and reliable measurements, but a detailed description of its intended use cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Tr0100, by Merck Group (5 mentions) Mak266, by Merck Group (4 mentions) Infinity cholesterol reagent, by Thermo Fisher Scientific (2 mentions) Mak045, by Merck Group (2 mentions) Mak044, by Merck Group (2 mentions) Microplate reader, by Thermo Fisher Scientific (2 mentions) Bca assay kit, by Beyotime (1 mentions) Mak309, by Merck Group (1 mentions) Mak187, by Merck Group (1 mentions) Khb3481, by Thermo Fisher Scientific (1 mentions) Khb3441, by Thermo Fisher Scientific (1 mentions) Kho0631, by Thermo Fisher Scientific (1 mentions) Tr15421, by Thermo Fisher Scientific (1 mentions) Mak041, by Merck Group (1 mentions) Onetouch ultraeasy, by LifeScan (1 mentions) Dcfda h2dcfda cellular ros assay kit, by Abcam (1 mentions) Mak085, by Merck Group (1 mentions) Tricorn high performance superose s 6 10 300gl column, by Cytiva (1 mentions) Mak175, by Merck Group (1 mentions) Mak052, by Merck Group (1 mentions)

Close spelling variants of this product

Cholesterol Quantitation Kit-MAK043 (6 citations) MAK043–1KT (3 citations) Cholesterol quantification kit (MAK043) (2 citations)

28 protocols using mak043

1

Quantitative Cholesterol Analysis in Cortex

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A cholesterol quantitation kit (MAK043, MilliporeSigma) was used to measure the concentration of cholesterol in the cortex. The samples prepared above were diluted at 1:10 with assay buffer and seven cholesterol standards (0.02 µg/µl to 0.14 µg/µl) were prepared. 100µl of standards and samples were pipetted into the appropriate wells of a clear 96 well plate. 50µl of reaction mix buffer including cholesterol probe, cholesterol enzyme mix and cholesterol esterase was added to each well. The plate was incubated at 37°C for 60 mins protected from light. The absorbance was read at 570nm and the concentration of cholesterol in the samples was calculated using the standard curve. The levels of cortical cholesterol were normalized to the wet weight of the starting tissue (µg/mg wet weight).
Lu F., Zhu J., Guo S., Wong B.J., Chehab F.F., Ferriero D.M, & Jiang X. (2018). Up-regulation of cholesterol 24-hydroxylase following hypoxia-ischemia in neonatal mouse brain. Pediatric research, 83(6), 1218-1227.
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2

Quantitative Cholesterol Analysis in Cortex

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A cholesterol quantitation kit (MAK043, MilliporeSigma) was used to measure the concentration of cholesterol in the cortex. The samples prepared above were diluted at 1:10 with assay buffer and seven cholesterol standards (0.02 µg/µl to 0.14 µg/µl) were prepared. 100µl of standards and samples were pipetted into the appropriate wells of a clear 96 well plate. 50µl of reaction mix buffer including cholesterol probe, cholesterol enzyme mix and cholesterol esterase was added to each well. The plate was incubated at 37°C for 60 mins protected from light. The absorbance was read at 570nm and the concentration of cholesterol in the samples was calculated using the standard curve. The levels of cortical cholesterol were normalized to the wet weight of the starting tissue (µg/mg wet weight).
Lu F., Zhu J., Guo S., Wong B.J., Chehab F.F., Ferriero D.M, & Jiang X. (2018). Up-regulation of cholesterol 24-hydroxylase following hypoxia-ischemia in neonatal mouse brain. Pediatric research, 83(6), 1218-1227.
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3

Cholesterol Quantification in Plasma and Liver

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Total cholesterol in undiluted plasma and bile was assessed with Infinity Cholesterol reagent (TR13421, Thermo Scientific, Waltham, MA) and concentrations determined by a standard curve. Liver cholesterol was extracted by homogenizing 50 mg of tissue in a TissueLyser with 1 mL chloroform:isopropanol:IGEPAL CA-630 (7:11:0.1). The organic phase was collected and dried at 50°C. Dried lipids were resuspended in 200μL cholesterol assay buffer (MAK043, Millipore Sigma, St. Louis, MO) and total cholesterol determined following manufacturer’s protocol.
To analyze lipoprotein size distributions, plasma was analyzed using a Superose 6 10-300GL column and size-exclusion fast protein liquid chromatography (FPLC). Fractions were assayed for total cholesterol and triglycerides as previously described [51 (link)].
Price T.R., Stapleton D.S., Schueler K.L., Norris M.K., Parks B.W., Yandell B.S., Churchill G.A., Holland W.L., Keller M.P, & Attie A.D. (2023). Lipidomic QTL in Diversity Outbred mice identifies a novel function for α/β hydrolase domain 2 (Abhd2) as an enzyme that metabolizes phosphatidylcholine and cardiolipin. PLOS Genetics, 19(7), e1010713.
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4

Quantifying Cholesterol in Plasma and Liver

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Total cholesterol in undiluted plasma and bile was assessed with Infinity Cholesterol reagent (TR13421, Thermo Scientific, Waltham, MA) and concentrations determined by a standard curve. Liver cholesterol was extracted by homogenizing 50 mg of tissue in a TissueLyser with 1 mL chloroform:isopropanol:IGEPAL CA-630 (7:11:0.1). The organic phase was collected and dried at 50°C. Dried lipids were resuspended in 200μL cholesterol assay buffer (MAK043, Millipore Sigma, St. Louis, MO) and total cholesterol determined following manufacturer’s protocol.
To analyze lipoprotein size distributions, plasma was analyzed using a Superose 6 10-300GL column and size-exclusion fast protein liquid chromatography (FPLC). Fractions were assayed for total cholesterol and triglycerides as previously described[47 (link)].
Price T.R., Stapleton D.S., Schueler K.L., Norris M.K., Parks B.W., Yandell B.S., Churchill G.A., Holland W.L., Keller M.P, & Attie A.D. (2023). Lipidomic QTL in Diversity Outbred mice identifies a novel function for α/β hydrolase domain 2 (Abhd2) as an enzyme that metabolizes phosphatidylcholine and cardiolipin. bioRxiv.
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5

Quantifying Macrophage Cholesterol Levels

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Macrophage foam cells (1 × 106) were washed with PBS and extracted with 200 μL of chloroform: isopropanol: NP-40 (7:11:0.1) in a microhomogenizer. After the cells were air dried and vacuumed, a cholesterol quantitation kit (MAK043, Sigma-Aldrich) was used to detect the total and free cholesterol; the concentration of cholesteryl ester = total cholesterol – free cholesterol. Protein content was detected using a BCA assay kit (Beyotime Biotechnology).
Lv J.J., Wang H., Cui H.Y., Liu Z.K., Zhang R.Y., Lu M., Li C., Yong Y.L., Liu M., Zhang H., Zhang T.J., Zhang K., Li G., Nan G., Zhang C., Guo S.P., Wang L., Chen Z.N, & Bian H. (2021). Blockade of Macrophage CD147 Protects Against Foam Cell Formation in Atherosclerosis. Frontiers in Cell and Developmental Biology, 8, 609090.
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6

Quantification of Lipid Metabolites

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TG and cholesterol levels were measured using kits (TR0100 and MAK043, respectively, Sigma) and total BA levels in the liver, intestine, plasma, and gallbladder were measured with a kit (MAK309, Sigma). Plasma metabolites were identified by gas chromatography-mass spectrometry (GC-MS) in the Metabolomics Center at UIUC.
Kim Y.C., Qi M., Dong X., Seok S., Sun H., Kemper B., Fu T, & Kemper J.K. (2023). Transgenic mice lacking FGF15/19-SHP phosphorylation display altered bile acids and gut bacteria, promoting nonalcoholic fatty liver disease. The Journal of Biological Chemistry, 299(8), 104946.
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7

Preparation and Quantification of ISCOM-like Saponin Adjuvant

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ISCOM-like saponin adjuvant was prepared as described41 . Final adjuvant concentration
was determined by cholesterol quantification (Sigma MAK043).
Escolano A., Gristick H.B., Abernathy M.E., Merkenschlager J., Gautam R., Oliveira T.Y., Pai J., West AP J.r., Barnes C.O., Cohen A.A., Wang H., Golijanin J., Yost D., Keeffe J.R., Wang Z., Zhao P., Yao K.H., Bauer J., Nogueira L., Gao H., Voll A.V., Montefiori D.C., Seaman M.S., Gazumyan A., Silva M., McGuire A.T., Stamatatos L., Irvine D.J., Wells L., Martin M.A., Bjorkman P.J, & Nussenzweig M.C. (2019). Immunization expands HIV-1 V3-glycan specific B-cells in mice and macaques. Nature, 570(7762), 468-473.
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8

Preparation and Quantification of ISCOM-like Saponin Adjuvant

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ISCOM-like saponin adjuvant was prepared as described41 . Final adjuvant concentration
was determined by cholesterol quantification (Sigma MAK043).
Escolano A., Gristick H.B., Abernathy M.E., Merkenschlager J., Gautam R., Oliveira T.Y., Pai J., West AP J.r., Barnes C.O., Cohen A.A., Wang H., Golijanin J., Yost D., Keeffe J.R., Wang Z., Zhao P., Yao K.H., Bauer J., Nogueira L., Gao H., Voll A.V., Montefiori D.C., Seaman M.S., Gazumyan A., Silva M., McGuire A.T., Stamatatos L., Irvine D.J., Wells L., Martin M.A., Bjorkman P.J, & Nussenzweig M.C. (2019). Immunization expands HIV-1 V3-glycan specific B-cells in mice and macaques. Nature, 570(7762), 468-473.
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9

Serum Lipid and Antioxidant Biomarkers

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Serum total cholesterol, triglyceride, and low-density lipoprotein-cholesterol (LDL-C) concentrations were determined colorimetrically with a spectrophotometer using triglyceride (TR0100), total cholesterol (MAK043), and LDL/VLDL (MAK045) kits from Sigma Aldrich, following the manufacturer's instructions. Serum and meat filtrates were used for measuring the activity of the glutathione peroxidase (GSH-Px) using a commercial assay kit (Sigma-Aldrich, G6137). Total antioxidant capacity (T-AOC) was determined using a commercial assay kit (Sigma-Aldrich, MAK187), and the activity of superoxide dismutase (SOD) enzyme was determined using a commercial assay kit (Sigma-Aldrich, 19160) following the instructions of the test kit.
Ibrahim D., Moustafa A., Shahin S.E., Sherief W.R., Abdallah K., Farag M.F., Nassan M.A, & Ibrahim S.M. (2021). Impact of Fermented or Enzymatically Fermented Dried Olive Pomace on Growth, Expression of Digestive Enzyme and Glucose Transporter Genes, Oxidative Stability of Frozen Meat, and Economic Efficiency of Broiler Chickens. Frontiers in Veterinary Science, 8, 644325.
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10

Quantifying Intracellular Cholesterol in AML-12 Cells

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Intracellular cholesterol levels in AML-12 cells were visualized using Filipin staining and quantitation was performed using Image J software. Four equal sized areas with 20 cells each were chosen in each image and the fluorescence intensity of each area was determined. These values were then used to calculate the average fluorescence of each image and then of control (set to 100%) and Ulk1 silenced group. Mouse liver total cholesterol was measured using commercial kit (MAK043, SIGMA-ALDRICH, United States).
Rajak S., Iannucci L.F., Zhou J., Anjum B., George N., Singh B.K., Ghosh S., Yen P.M, & Sinha R.A. (2020). Loss of ULK1 Attenuates Cholesterogenic Gene Expression in Mammalian Hepatic Cells. Frontiers in Cell and Developmental Biology, 8, 523550.
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