Chod pap method

Manufactured by Roche

Sourced in Germany

The CHOD-PAP method is a laboratory technique used for the quantitative determination of cholesterol levels in biological samples. It is a colorimetric assay that relies on the enzymatic conversion of cholesterol by cholesterol oxidase (CHOD) and the subsequent reaction with 4-aminophenazone (PAP) to produce a colored complex, which is measured photometrically. This method provides a reliable and standardized way to measure cholesterol concentrations in clinical and research settings.

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Lab products found in correlation

Gpo tinder, by Merck Group (3 mentions) Gpo trinder, by Merck Group (2 mentions) Serum stt 2 advance tubes, by BD (2 mentions) Glucose hk cp, by Horiba (2 mentions) Nitroprusside, by Merck Group (1 mentions) Quantimet 570, by Leica (1 mentions) Serum separator tube, by BD (1 mentions)

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CHOD-PAP (72 citations)

9 protocols using chod pap method

Lipid Profile Changes After Exercise

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Twelve-hour fasting venous blood samples were drawn at baseline, week 4 and at two follow up days in week 8. The first follow up sample was collected 43 h (median: range 19–72 h) and the second follow up sample 117 h (70–118 h) after an exercise training. Blood samples were centrifuged at 1300x g for 15 min at 21 °C to collect serum samples that were stored at − 80 °C until analyzed at the end of the study. Serum TC (CHOD-PAP method; Roche Diagnostic, Mannheim, Germany), HDL-C (CHOD-PAP method; Roche Diagnostic, Mannheim, Germany), and TG concentrations were analyzed with enzymatic methods (GPO-Tinder; Sigma-Aldrich Corp., St. Louis, MO, USA).

Mashnafi S., Plat J., Mensink R.P., Joris P.J., Kleinloog J.P, & Baumgartner S. (2021). Effects of an 8-week aerobic exercise program on plasma markers for cholesterol absorption and synthesis in older overweight and obese men. Lipids in Health and Disease, 20, 112.

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Plasma Lipid and HDL Functionality Assay

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In the samples collected at baseline and after 24 h, plasma TC (CHOD-PAP method; Roche Diagnostic, Mannheim, Germany), HDL-C (CHOD-PAP method; Roche Diagnostic, Mannheim, Germany), and TG concentrations were analyzed enzymatically (GPO-Tinder; Sigma-Aldrich Corp., St. Louis, MO, USA). In these samples, plasma LDL-C concentrations were calculated using the Friedewald equation [22 (link)]. HDL functionality, defined as the capacity of radioactive cholesterol efflux from cultured J774 macrophages, using liquid scintillation counting was determined as described elsewhere [23 (link)]. Markers for inflammation were analyzed in the plasma samples collected at all time points as described [13 (link)].

Mashnafi S., Baumgartner S., Mensink R.P., Perlee D., van Vught L.A., Lütjohann D, & Plat J. (2023). A Transient Inflammatory Response Induced by Lipopolysaccharide Infusion Lowers Markers of Endogenous Cholesterol and Bile Acid Synthesis in Healthy Normocholesterolemic Young Men. Biomedicines, 11(1), 126.

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Automated Lipid Profile Analysis

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Serum total cholesterol (TC) (CHOD-PAP method; Roche Diagnostic, Mannheim, Germany), high-density lipoprotein cholesterol (HDL-C) (CHOD-PAP method; Roche Diagnostic, Mannheim, Germany), and triglyceride (TG) concentrations—corrected for glycerol levels—were analyzed enzymatically (GPO-Tinder; Sigma-Aldrich Corp., St. Louis, MO, USA). Serum low-density lipoprotein cholesterol (LDL-C) concentrations were calculated using the Friedewald equation [18 (link)].

Mashnafi S., Plat J., Mensink R.P., Joris P.J., Kusters Y.H., Houben A.J., Stehouwer C.D., Schalkwijk C.G, & Baumgartner S. (2022). Effects of Diet-Induced Weight Loss on Plasma Markers for Cholesterol Absorption and Synthesis: Secondary Analysis of a Randomized Trial in Abdominally Obese Men. Nutrients, 14(8), 1546.

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Enzymatic Cholesterol and Triglyceride Assay

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Concentrations of serum total cholesterol (CHOD-PAP method; Roche Diagnostics Systems, Hoffmann-La Roche) and TAG with correction for free glycerol (GPO-Trinder; Sigma Diagnostics) were determined enzymatically in fasting serum samples.

De Smet E., Mensink R.P., Boekschoten M.V., de Ridder R., Germeraad W.T., Wolfs T.G, & Plat J. (2015). An acute intake of plant stanol esters alters immune-related pathways in the jejunum of healthy volunteers. The British journal of nutrition, 113(5).

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Circadian Analysis of Metabolic Parameters

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Rats from each group were sacrificed at ZT 2:00, 6:00, 10:00, 14:00, 18:00 and 22:00 h. The liver tissues were immediately frozen in liquid nitrogen and stored in RNAlater (cat. no. AM7020; Ambion; Thermo Fisher Scientific, Inc., Waltham, MA, USA) at −80°C. Blood samples were centrifuged at 6,391 × g for 10 min at 4°C. Samples were sent to the Department of Laboratory Medicine, PUMCH (Beijing, China). The following measurements were performed with a Hitachi Modular P800 analyzer (Hitachi, Ltd., Tokyo, Japan): Serum total cholesterol (TC) (CHOD-PAP method; Roche Diagnostics GmbH, Mannheim, Germany) (18 (link)), serum TGs (GP0-PAP method; Roche Diagnostics GmbH), and HDL-C (Roche HDL-C Plus 2nd generation kit; Roche Diagnostics GmbH). LDL-C was calculated using the Friedewald formula (19 (link)). Albumin was measured using the bromopotassium phenol green method (20 (link)) and serum creatinine was measured using a sarcosine oxidase method (21 (link)). Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activity was assessed with an automatic biochemical analyzer. Furthermore, GLU levels were measured using the hexokinase method (22 (link)).

Chen P., Zhang R., Mou L., Li X., Qin Y, & Li X. (2018). An impaired hepatic clock system effects lipid metabolism in rats with nephropathy. International Journal of Molecular Medicine, 42(5), 2720-2736.

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Quantification of Carotid Artery Intima-Media

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Four weeks after ligation, mice were sacrificed. The arterial tree was perfused with PBS containing 0.1 mg/ml nitroprusside (Sigma, St Louis, MO) and subsequently with 1% paraformaldehyde via a catheter inserted into the left cardiac ventricle. Right carotid artery cross-sections (4 μm thick) were cut at 100 μm intervals. At each level of 100 μm, a cross-section was stained with Lawson solution and used to determine intima, lumen and total vessel area as well as elastic lamina fragmentation. The relative intimal collagen content, i.e. the percentage of total intima area that stained positive for Sirius red, was determined under a microscope coupled to a computerized morphometry system (Quantimet 570, Leica). Morphometric analyses were performed by a blinded investigator (LB, intra-observer variability was < 10%) using a computerized morphometry system (Quantimet 570, Leica). Serum cholesterol level was determined with the CHOD-PAP method (Roche Diagnostics).

Donners M.M., Bai L., Lutgens S.P., Wijnands E., Johnson J., Schurgers L.J., Liu C.L., Daemen M.J., Cleutjens K.B., Shi G.P., Biessen E.A, & Heeneman S. (2016). Cathepsin K Deficiency Prevents the Aggravated Vascular Remodeling Response to Flow Cessation in ApoE-/- Mice. PLoS ONE, 11(9), e0162595.

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Fasting Biomarker Assessment Protocol

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Fasting blood samples were drawn from the antecubital vein by venipuncture (BL, MT, and FU-A) or from an intravenous catheter (FUeB). Blood in serum separator tubes (Becton, Dickinson and Company, Franklin Lakes, NY, USA) was allowed to clot for at least 60 min at room temperature before samples were centrifuged (15 min at 1500 g at room temperature), distributed in aliquots, snap frozen, and stored at À80 C until analysis at the end of the study. Serum was used for the analysis of C-reactive protein (CRP; immunoturbidimetric assay, Horiba ABX, Montpellier, France), TAG with correction for free glycerol (Trinder; SigmaeAldrich Corp., St. Louis, MO, USA), TC, and high-density lipoprotein (HDL)-cholesterol (CHOD-PAP method; Roche Diagnostics System, Mannheim, Germany), and isoflavone concentrations (i.e., daidzein, genistein, and equol; LC-MS/MS technique by LGC Limited, analyzed at LGC Limited, Fordham, UK) [24] . Serum concentrations of daidzein and genistein were used to measure compliance, while serum equol concentrations were determined to differentiate between equolproducers and non-producers [25] . Finally, serum low-density lipoprotein (LDL)-cholesterol concentrations were calculated using the Friedewald formula [26] .

Tischmann L., Adam T.C., Mensink R.P, & Joris P.J. (2022). Longer-term soy nut consumption improves vascular function and cardiometabolic risk markers in older adults: Results of a randomized, controlled cross-over trial. Clinical nutrition (Edinburgh, Scotland), 41(5).

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Plasma Glucose and Lipid Biomarkers

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Plasma prepared from blood collected in sodium fluoride (NaF) and Na₂EDTA (Becton Dickinson, Erembodegem, Belgium) tubes were used for the determination of plasma glucose concentrations (Glucose HK CP, Horiba ABX, Montpellier, France) at all time points. In addition, blood was sampled in serum STT-II advance tubes (Becton Dickinson) for the determination of serum triacylglycerol (TAG; GPO Trinder, Sigma-Aldrich Corp., St. Louis, MO, USA) concentrations with correction for free glycerol at T0, T60, T120, and T180. In addition, serum total (CHOD-PAP method; Roche Diagnostic Systems), high-density lipoprotein cholesterol (HDL-cholesterol; precipitation method (phosphotungstate precipitant); Roche Diagnostics, Mannheim, Germany) and high-sensitivity C-reactive protein (hsCRP; CRP CP, Horiba ABX, Montpellier, France) concentrations at T0 were measured. Fasting serum low-density lipoprotein (LDL)-cholesterol concentrations were calculated using the Friedewald formula [13 (link)]. More details regarding the biochemical analyses can be found in our previous study [12 (link)].

Smeets E.T., Mensink R.P., Hoeks J., de Vogel-Van den Bosch J., Hageman R.J, & Joris P.J. (2020). Effects of Beetroot Powder with or without L-Arginine on Postprandial Vascular Endothelial Function: Results of a Randomized Controlled Trial with Abdominally Obese Men. Nutrients, 12(11), 3520.

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Blood Sample Collection and Biochemical Analyses

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Blood was collected in serum STT-II advance tubes (Becton Dickinson). After a minimum of 30 min clotting time at room temperature, tubes were centrifuged at 1300 g for 10 min at 21°C to obtain serum. In addition, on day 0, blood was collected in sodium fluoride (NaF) tubes (Becton Dickinson). Directly after sampling, these NaF tubes were centrifuged at 1300 g for 10 min at 4 °C to obtain plasma. Subsequently, serum and plasma were portioned into aliquots, frozen in liquid nitrogen, and stored at -80°C until analyses.
Brain-derived neurotrophic factor (BDNF) was measured in serum samples of day 0 and 27 of each period by an enzyme-linked immunosorbent assay (Duo Kit ELISA, R & D Systems, Minneapolis, MN, United States) according to manufacturer's instructions. Fasting TAG (GPO Trinder, Sigma-Aldrich Corp., St. Louis, MO, United States), total cholesterol (CHOD-PAP method; Roche Diagnostic Systems), and HDL cholesterol concentrations (precipitation method; Roche Diagnostics, Mannheim, Germany) were measured in serum samples at baseline. LDL cholesterol was calculated using the Friedewald equation [Citation15] . Fasting glucose concentrations (Glucose HK CP, Horiba ABX, Montpellier, France) were measured in NaF plasma samples at baseline.

Gravesteijn E., Adam J.J., Mensink R.P., Winkens B, & Plat J. (2023). Effects of the egg protein hydrolysate NWT-03 on cognitive function in men and women with the metabolic syndrome: a randomized, double-blind, placebo-controlled study. Nutritional neuroscience, 26(12).

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