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Mpc900

Manufactured by R&D Systems
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Sourced in United States

The MPC900 is a multi-parameter cell culture incubator that maintains optimal environmental conditions for cell culture. It features precise temperature, CO2, and humidity control to support the growth and maintenance of various cell types.

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Lab products found in correlation

Dpc900, by R&D Systems (3 mentions) Total cholesterol reagents tr13421, by Thermo Fisher Scientific (2 mentions) Ab230932, by Abcam (2 mentions) Mak052, by Merck Group (1 mentions) Sb 1010 225, by Thermo Fisher Scientific (1 mentions) Tr213, by Randox (1 mentions) Ab241035, by Abcam (1 mentions) Ab238260, by Abcam (1 mentions)

9 protocols using mpc900

1

Plasma Cholesterol and Pcsk9 Quantification

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After harvesting mice, plasma samples were aliquoted and stored at −80 °C. Plasma levels of cholesterol were measured via a colorimetric assay according to the manufacturer’s instructions (Thermo- Scientific Total Cholesterol Reagents #TR13421). Plasma Pcsk9 protein levels were quantified by ELISA according to the manufacturer’s instructions (R&D Systems #MPC900).
Moghadam F., LeGraw R., Velazquez J.J., Yeo N.C., Xu C., Park J., Chavez A., Ebrahimkhani M.R, & Kiani S. (2020). Synthetic immunomodulation with a CRISPR super-repressor in vivo. Nature cell biology, 22(9), 1143-1154.
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2

Plasma PCSK9 Quantification by ELISA

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Peripheral blood was collected in EDTA-coated capillary tubes from vena saphena during the course of the study and by cardiac puncture at the time of termination. Samples were kept on ice for up to 2 hours prior to extraction of plasma by centrifugation at 10,000 rpm for 20 min at 4oC. Plasma was stored at −80°C until the samples were analyzed. Plasma human PCSK9 and mouse Pcsk9 levels were determined with a standard ELISA kit (DPC900 and MPC900; R&D Systems, Minneapolis, MN, USA) according to the manufacturer´s instructions. Prior to the assay, plasma samples were diluted 1:800 and 1:1000 for human PCSK9 and mouse Pcsk9, respectively.
Akcakaya P., Bobbin M.L., Guo J.A., Malagon-Lopez J., Clement K., Garcia S.P., Fellows M.D., Porritt M.J., Firth M.A., Carreras A., Baccega T., Seeliger F., Bjursell M., Tsai S.Q., Nguyen N.T., Nitsch R., Mayr L.M., Pinello L., Bohlooly-Y M., Aryee M.J., Maresca M, & Joung J.K. (2018). In vivo CRISPR editing with no detectable genome-wide off-target mutations. Nature, 561(7723), 416-419.
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3

Quantification of PCSK9, LDL-C, and Liver Enzymes

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To quantify PCSK9, plasma from treated mice and supernatants from primary mouse hepatocytes were thawed and diluted 1:200 and 1:2, respectively. Dilutions were then loaded on a commercial pre-spotted ELISA kit according to the manufacturer’s instructions (R&D Systems, MPC900). Similarly, absorbance assays were used to quantify the levels of LDL-C (P/N 00018256040, Werfen), ALT (P/N 00018257440, Werfen), AST (P/N 00018257540, Werfen), LDH (P/N 00018258240, Werfen) and albumin (P/N 00 18250040), following the manufacturer’s instructions.
Cappelluti M.A., Mollica Poeta V., Valsoni S., Quarato P., Merlin S., Merelli I, & Lombardo A. (2024). Durable and efficient gene silencing in vivo by hit-and-run epigenome editing. Nature, 627(8003), 416-423.
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4

Serum Biomarker Quantification Protocol

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After harvest, serum was stored in one-time use aliquots at −80 °C. Total cholesterol and LDL cholesterol levels were measured from serum via a colorimetric assay according to the manufacturer’s instructions (ThermoScientific Total Cholesterol Reagents #TR13421 and WakoChemical LDL Cholesterol #993–00404). ALT secretion was measured from serum via a colorimetric assay according to the manufacturer’s instructions (Sigma-Aldrich #MAK052). Pcsk9 serum protein levels were quantified by ELISA with a standard curve according to the manufacturer’s instructions (R&D Systems #MPC900).
Thakore P.I., Kwon J.B., Nelson C.E., Rouse D.C., Gemberling M.P., Oliver M.L, & Gersbach C.A. (2018). RNA-guided transcriptional silencing in vivo with S. aureus CRISPR-Cas9 repressors. Nature Communications, 9, 1674.
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5

Multiparametric Serum Lipoprotein Analysis

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Sera were analyzed by a colorimetric assay for total cholesterol (SB-1010–225, Fisher Scientific, Hampton NH) and by ELISAs for PCSK9 (MPC900, R&D Systems, Minneapolis MN) and APOB (ab230932, abcam, Cambridge UK). Serum lipoprotein fractionation assays were performed at the University of Cincinnati Mouse Metabolic Phenotyping Center. Sera were pooled from 5 mice for each genotype and fractionated by fast liquid protein chromatography (FPLC) into 50 fractions. Cholesterol (NC9343696, Fisher, Hampton NH) and triglyceride (TR213, Randox Laboratories, Crumlin UK) content in each fraction were determined using a microliter plate enzyme-based assay. Liver function tests were performed at the University of Michigan In-Vivo Animal Core (IVAC) with sera collected from individual mice using a Liasys analyzer (AMS Alliance).
Tang V.T., McCormick J., Xu B., Wang Y., Fang H., Wang X., Siemieniak D., Khoriaty R., Emmer B.T., Chen X.W, & Ginsburg D. (2022). Hepatic inactivation of murine Surf4 results in marked reduction in plasma cholesterol. eLife, 11, e82269.
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6

Mouse Pcsk9 and Human PCSK9 Plasma Levels

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Over the course of the study, mouse blood was collected into EDTA-coated tubes from the vena saphena (pre-injection) and from cardiac puncture (during termination). Tubes were kept on ice and plasma was isolated by 20 min centrifugation at 12,000 × g and 4 °C. For assessment of mouse Pcsk9 and human PCSK9 plasma levels, the samples were diluted 1:1000 and 1:800, respectively, and measured with standard ELISA kits (DPC900 and MPC900; R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instructions.
Bestas B., Wimberger S., Degtev D., Madsen A., Rottner A.K., Karlsson F., Naumenko S., Callahan M., Touza J.L., Francescatto M., Möller C.I., Badertscher L., Li S., Cerboni S., Selfjord N., Ericson E., Gordon E., Firth M., Chylinski K., Taheri-Ghahfarokhi A., Bohlooly-Y M., Snowden M., Pangalos M., Nuttall B., Akcakaya P., Sienski G, & Maresca M. (2023). A Type II-B Cas9 nuclease with minimized off-targets and reduced chromosomal translocations in vivo. Nature Communications, 14, 5474.
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7

Plasma PCSK9 Quantification by ELISA

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Peripheral blood was collected in EDTA-coated capillary tubes from vena saphena during the course of the study and by cardiac puncture at the time of termination. Samples were kept on ice for up to 2 hours prior to extraction of plasma by centrifugation at 10,000 rpm for 20 min at 4oC. Plasma was stored at −80°C until the samples were analyzed. Plasma human PCSK9 and mouse Pcsk9 levels were determined with a standard ELISA kit (DPC900 and MPC900; R&D Systems, Minneapolis, MN, USA) according to the manufacturer´s instructions. Prior to the assay, plasma samples were diluted 1:800 and 1:1000 for human PCSK9 and mouse Pcsk9, respectively.
Akcakaya P., Bobbin M.L., Guo J.A., Malagon-Lopez J., Clement K., Garcia S.P., Fellows M.D., Porritt M.J., Firth M.A., Carreras A., Baccega T., Seeliger F., Bjursell M., Tsai S.Q., Nguyen N.T., Nitsch R., Mayr L.M., Pinello L., Bohlooly-Y M., Aryee M.J., Maresca M, & Joung J.K. (2018). In vivo CRISPR editing with no detectable genome-wide off-target mutations. Nature, 561(7723), 416-419.
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8

Plasma Cholesterol and Pcsk9 Quantification

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After harvesting mice, plasma samples were aliquoted and stored at −80 °C. Plasma levels of cholesterol were measured via a colorimetric assay according to the manufacturer’s instructions (Thermo- Scientific Total Cholesterol Reagents #TR13421). Plasma Pcsk9 protein levels were quantified by ELISA according to the manufacturer’s instructions (R&D Systems #MPC900).
Moghadam F., LeGraw R., Velazquez J.J., Yeo N.C., Xu C., Park J., Chavez A., Ebrahimkhani M.R, & Kiani S. (2020). Synthetic immunomodulation with a CRISPR super-repressor in vivo. Nature cell biology, 22(9), 1143-1154.
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9

Plasma Lipid and Enzyme Profiling

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Circulating levels of PCSK9 (MPC900, R&D), ApoB (ab230932, Abcam) and ApoA1 (ab238260, Abcam) levels were measured using a commercially available mouse ELISA kits. Plasma ALT was measured using a commercially available colorimetric assay (ab241035, Abcam). All assays were performed according to the manufacturer’s instructions.
Platko K., Lebeau P.F., Nederveen J.P., Byun J.H., MacDonald M.E., Bourgeois J.M., Tarnopolsky M.A, & Austin R.C. (2023). A Metabolic Enhancer Protects against Diet-Induced Obesity and Liver Steatosis and Corrects a Pro-Atherogenic Serum Profile in Mice. Nutrients, 15(10), 2410.
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