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Tr13421

Manufactured by Thermo Fisher Scientific
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The TR13421 is a laboratory centrifuge designed for general-purpose applications. It features a maximum speed of 6,000 rpm and can accommodate rotor sizes up to 200 mL. The centrifuge provides reliable performance and is suitable for a variety of sample processing tasks.

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Lab products found in correlation

Tr22421, by Thermo Fisher Scientific (12 mentions) Nefa hr 2, by Fujifilm (3 mentions) Tr71121, by Thermo Fisher Scientific (2 mentions) Tr70121, by Thermo Fisher Scientific (2 mentions) Tr15421, by Thermo Fisher Scientific (2 mentions) Onetouch ultraeasy, by Johnson & Johnson (2 mentions) Triglyceride reagent, by Thermo Fisher Scientific (1 mentions) Total cholesterol reagent, by Thermo Fisher Scientific (1 mentions) Mak041, by Merck Group (1 mentions) Ezrmcp2 21k, by Merck Group (1 mentions) Ezrmi 13k, by Merck Group (1 mentions) P 407 pluronic f127, by Merck Group (1 mentions)

14 protocols using tr13421

1

Quantifying Atherosclerosis and Inflammatory Markers

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Secreted IL-1β and TNF-α from peritoneal macrophage culture media were measured using ELISA assays from R&D Systems (MLB00B) and BD Biosciences (560478). Cholesterol, triglycerides, and glucose were assayed in serum following a 6-hour fast per manufacturer’s protocols (Thermo Scientific TR13421, TR22421, and TR15408). Proteasome activity was assayed in macrophages using the Proteasome-Glo luminescent kit per manufacturer’s protocol (Promega G8660). For assessment of transcripts, extraction of total RNA, reverse transcription, and quantitative RT-PCR was performed as described (59 (link)). Sequences for the oligonucleotide primers used were designed using the qPrimerDepot database- http://primerdepot.nci.nih.gov. Assays were performed in duplicate and normalized to amplified mRNA for the ribosomal protein 36B4.
The en face and aortic root cross-section techniques previously described (60 (link)) were used to quantify atherosclerosis throughout the aorta as well as at the vessel origin. En face regions analyzed included the aortic arch, thoracic aorta, and abdominal aorta (spanning from the aortic valve to the bifurcation of the iliac arteries).
Sergin I., Bhattacharya S., Emanuel R., Esen E., Stokes C.J., Evans T.D., Arif B., Curci J.A, & Razani B. (2016). p62-enriched inclusion bodies in macrophages protect against atherosclerosis. Science signaling, 9(409), ra2.
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2

Blood Lipid Measurement Protocol

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To analyze blood lipid levels, triglyceride and cholesterol levels were measured using triglyceride reagent (TR22421; Thermo Scientific) and total cholesterol reagent (TR13421; Thermo Scientific) according to the manufacturer’s instructions.
Yamada T., Murata D., Kleiner D.E., Anders R., Rosenberg A.Z., Kaplan J., Hamilton J.P., Aghajan M., Levi M., Wang N.Y., Dawson T.M., Yanagawa T., Powers A.F., Iijima M, & Sesaki H. (2022). Prevention and regression of megamitochondria and steatosis by blocking mitochondrial fusion in the liver. iScience, 25(4), 103996.
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3

Quantification of Metabolic Biomarkers in Mice

Cited 2 times
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Mouse plasma acylcarnitines were measured after derivatization to butylesters according to a standardized protocol (Violante et al. 2019 (link)). Tissue acylcarnitines were measured in freeze-dried tissue samples (approximately 50 mg of liver) after derivatization to propylesters essentially as described (van Vlies et al. 2005 (link); Ranea-Robles et al. 2020 (link)). Organic acids from mouse urine were analyzed by GC-MS as previously described (Ranea-Robles et al. 2021b (link)).
Plasma bile acids were analyzed by LC-negative ion electrospray MS/MS as previously described (Bootsma et al. 1999 (link); Ferdinandusse et al. 2005 (link)).
Total glycerol (i.e. free glycerol and triglycerides, TR22421, Thermo Scientific), total cholesterol (TR13421, Thermo Scientific), non-esterified fatty acids (NEFA-HR(2), Wako), β-hydroxybutyrate (β-OHB, MAK-041, Sigma Aldrich), alanine aminotransferase (ALT, TR71121, Thermo Scientific) and aspartate aminotransferase (AST, TR70121, Thermo Scientific) plasma levels were measured using commercial kits following manufacturer instructions.
Ranea-Robles P., Chen H., Stauffer B., Yu C., Bhattacharya D., Friedman S.L., Puchowicz M, & Houten S.M. (2021). The peroxisomal transporter ABCD3 plays a major role in hepatic dicarboxylic fatty acid metabolism and lipid homeostasis. Journal of inherited metabolic disease, 44(6), 1419-1433.
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4

Serum Lipid Profiling in Fasted Mice

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Mice were fasted for 5 hours in the morning before tail vein bleeding to collect serum. Colorimetric kit assays were used to measure glucose (TR14598, Thermo Fisher Scientific), triglycerides (TR22421, Thermo Fisher Scientific), free fatty acids (NC9517308–12, Wako Diagnostics), and cholesterol (TR13421, Thermo Fisher Scientific).
Evans T.D., Zhang X., Jeong S.J., He A., Song E., Bhattacharya S., Holloway K.B., Lodhi I.J, & Razani B. (2019). TFEB drives PGC-1α expression in adipocytes to protect against diet-induced metabolic dysfunction. Science signaling, 12(606), eaau2281.
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5

Comprehensive Metabolic Profiling Protocol

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Plasma fructosamine levels were measured with a colorimetric kit (K450-100, BioVision Inc). Plasma insulin and C-peptide were measured with mouse/rat ELISAs (EZRMI-13K and EZRMCP2-21K, respectively, EMD Millipore). Plasma NEFAs were measured by enzymatic colorimetric assay (NEFA-HR(2), FUJIFILM Wako). Plasma triacylglycerol (TAG) and cholesterol were measured with colorimetric assays (TR22421 and TR13421, respectively, ThermoFisher Scientific). Plasma ALT and AST concentrations were measured with kinetic spectrophotometric assays (A524-150 and A559-150, respectively, Teco Diagnostics).
Kamm D.R., Pyles K.D., Sharpe M.C., Healy L.N., Colca J.R, & McCommis K.S. (2021). Novel insulin sensitizer MSDC-0602K improves insulinemia and fatty liver disease in mice, alone and in combination with liraglutide. The Journal of Biological Chemistry, 296, 100807.
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6

P-407 Hyperlipidemia Induction Protocol

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P‐407 (Pluronic F‐127, Cat. no. P2443; Sigma‐Aldrich, Dorset, UK) was dissolved overnight in cooled sterile phosphate‐buffered saline (PBS). Mice were injected intraperitoneally (i.p.) with 200 μl of P‐407 solution (10, 5 or 2·5 mg equivalent to 0·5, 0·25 and 0·125 g/kg, respectively) or PBS every other day. According to the experimental design blood samples were collected into heparin tubes at different time‐points. In P‐407‐treated animals, blood collection was performed 24 h following drug administration. CHOL and TG levels were measured by colorimetric assay using the cholesterol and triglyceride infinity reagent (TR13421 and TR22421, respectively; Thermo‐Scientific, Middletown, VA, USA).
Saja M.F., Cook H.T., Ruseva M.M., Szajna M., Pickering M.C., Woollard K.J, & Botto M. (2018). A triglyceride‐rich lipoprotein environment exacerbates renal injury in the accelerated nephrotoxic nephritis model. Clinical and Experimental Immunology, 192(3), 337-347.
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7

Quantifying Liver Cholesterol Levels

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To quantify cholesterol levels, liver samples (50-100 mg) were homogenized in 2 ml Folch (chloroform/methanol, 2:1, v/v) with a polytron homogenizer. The organic phase was separated with 1 mL of water and centrifugation and then dried under nitrogen. Samples were reconstituted in isopropranol:Triton-X100 (9:1 v/v) and aliquots subjected to colorimetric enzymatic assays for total cholesterol (ThermoFisher, TR13421).
Osinski V., Bauknight D.K., Dasa S.S., Harms M.J., Kroon T., Marshall M.A., Garmey J.C., Nguyen A.T., Hartman J., Upadhye A., Srikakulapu P., Zhou A., O'Mahony G., Klibanov A.L., Kelly K.A., Boucher J, & McNamara C.A. (2020). In vivo liposomal delivery of PPARα/γ dual agonist tesaglitazar in a model of obesity enriches macrophage targeting and limits liver and kidney drug effects. Theranostics, 10(2), 585-601.
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8

Plasma Lipid Profiling in Mice

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Plasma samples were obtained by cardiac puncture from mice fasted overnight. Total plasma cholesterol and triglyceride levels were determined by commercially available enzymatic kits (TR13421 and TR22421, Thermo Scientific, Waltham, MA) following manufacturer`s instructions. For lipoprotein profile analysis, plasma samples were prepared by using Microvette 500K3E EDTA-columns (Sarstedt, Germany) following the same procedure as described for serum. Plasma lipoproteins were fractionated by fast-protein liquid chromatography (FPLC) using a Superose six column (10/300 GL). Cholesterol and triglyceride content of all fractions was measured by using the same method as described for serum lipid levels.
Xian X., Ding Y., Dieckmann M., Zhou L., Plattner F., Liu M., Parks J.S., Hammer R.E., Boucher P., Tsai S, & Herz J. (2017). LRP1 integrates murine macrophage cholesterol homeostasis and inflammatory responses in atherosclerosis. eLife, 6, e29292.
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9

Glucose and Insulin Tolerance Tests

Cited 1 time
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GTT and ITT were performed as described previously (Xu et al., 2019 (link)). In brief, for GTT, 6-h fasted mice were injected i.p. with D-glucose (1 g·kg−1), and blood samples were collected and measured at 0, 30, 60, 90, and 120 min after glucose injection. For ITT, 4-h fasted mice were given insulin (1 U·kg−1), and blood samples were collected and measured at the same time points as GTT with a glucometer (OneTouch UltraEasy, Johnson & Johnson). Body composition of mice was assessed by NMR (MiniSpec LF90II TD-NMR Analyzer). Oxygen consumption (VO2), carbon dioxide production (VCO2), food intake, and heat production were monitored with CLAMS (Columbus Instruments). At the end of HFD administration, mice were anaesthetised with ketamine (80 mg·kg−1) and xylazine (5 mg·kg−1), given i.p.. Mice were fasted for 12–14 h before blood was collected. Individual kits were purchased from Thermo Scientific for measuring serum concentrations of cholesterol, triglyceride, and glucose (#TR15421, TR13421, and TR22421; Thermo Scientific). The content of hepatic triglyceride was determined using tissue lipids extracted by a mixture of chloroform and methanol as previously described (Xu et al., 2019 (link)).
Yang Q., Ma Q., Xu J., Liu Z., Mao X., Zhou Y., Cai Y., Da Q., Hong M., Weintraub N.L., Fulton D.J., Belin de Chantemèle E.J, & Huo Y. (2021). Endothelial AMPKα1/PRKAA1 exacerbates inflammation in HFD‐fed mice. British Journal of Pharmacology, 179(8), 1661-1678.
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10

Quantifying Cellular Energy, Lipids, and Cholesterol

Cited 1 time
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ATP was measured with a commercially available kit (FLAA, Sigma) using bioluminescence at 560 nm, as described previously (Sentelle et al., 2012 (link)). TAG and cholesterol were measured using commercially available kits TR22421and TR13421 (ThermoFisher), as described in the manufacturer’s protocol.
Chakraborty P., Vaena S.G., Thyagarajan K., Chatterjee S., Al-Khami A., Selvam S.P., Nguyen H., Kang I., Wyatt M.W., Baliga U., Hedley Z., Ngang R.N., Guo B., Beeson G.C., Husain S., Paulos C.M., Beeson C.C., Zilliox M.J., Hill E.G., Mehrotra M., Yu X.Z., Ogretmen B, & Mehrotra S. (2019). Pro-Survival Lipid Sphingosine-1-Phosphate Metabolically Programs T Cells to Limit Anti-tumor Activity. Cell reports, 28(7), 1879-1893.e7.
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