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Mouse proprotein convertase 9 pcsk9 quantikine elisa kit

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The Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit is a quantitative sandwich enzyme immunoassay designed to measure mouse PCSK9 levels in cell culture supernates, serum, and plasma. It utilizes a microplate pre-coated with a monoclonal antibody specific for mouse PCSK9.

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Lab products found in correlation

Infinity cholesterol reagent, by Thermo Fisher Scientific (3 mentions) Au5400, by Olympus (2 mentions) Hdl and ldl vldl cholesterol assay kit, by Abcam (1 mentions) Infinity triglycerides reagent, by Thermo Fisher Scientific (1 mentions) Human proprotein convertase 9 pcsk9 quantikine elisa kit, by R&D Systems (1 mentions) Cox fluorescent activity assay kit, by Cayman Chemical (1 mentions) Sandwich type elisa, by Thermo Fisher Scientific (1 mentions) Percoll, by Merck Group (1 mentions)

Close spelling variants of this product

PCSK9 Quantikine ELISA Kit PCSK9 ELISA kit Quantikine ELISA kit Quantikine ELISA Mouse ELISA kits PCSK9 Quantikine kits Quantikines ELISAs

9 protocols using mouse proprotein convertase 9 pcsk9 quantikine elisa kit

1

Quantifying Mouse Liver Metabolism and Lipids

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Following tissue harvesting as described above, flash-frozen mouse liver samples were embedded in O.C.T. compound (Tissue Tek, Cat # 4583), snap-frozen, and stored at −80°C prior to processing. Frozen tissues were cryosectioned at 4-micron in thickness and stained with Oil Red O following manufacturer’s recommended protocol. Liver histology was assessed by H&E staining sections of 10% neutral buffer formalin fixed liver sections.
Serum levels of Pcsk9 were determined by ELISA using the Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit (MPC-900, R&D Systems), following the manufacturer’s instructions. Total cholesterol levels were measured using the Infinity Cholesterol Reagent (Thermo Fisher) per the manufacturer’s instructions. Serum ALT, AST, albumin and total bilirubin were measured by an Olympus AU5400 (IDEXX Memphis, TN).
Ran F.A., Cong L., Yan W.X., Scott D.A., Gootenberg J.S., Kriz A.J., Zetsche B., Shalem O., Wu X., Makarova K.S., Koonin E., Sharp P.A, & Zhang F. (2015). In vivo genome editing using Staphylococcus aureus Cas9. Nature, 520(7546), 186-191.
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2

Quantifying Mouse Liver Metabolism and Lipids

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Following tissue harvesting as described above, flash-frozen mouse liver samples were embedded in O.C.T. compound (Tissue Tek, Cat # 4583), snap-frozen, and stored at −80°C prior to processing. Frozen tissues were cryosectioned at 4-micron in thickness and stained with Oil Red O following manufacturer’s recommended protocol. Liver histology was assessed by H&E staining sections of 10% neutral buffer formalin fixed liver sections.
Serum levels of Pcsk9 were determined by ELISA using the Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit (MPC-900, R&D Systems), following the manufacturer’s instructions. Total cholesterol levels were measured using the Infinity Cholesterol Reagent (Thermo Fisher) per the manufacturer’s instructions. Serum ALT, AST, albumin and total bilirubin were measured by an Olympus AU5400 (IDEXX Memphis, TN).
Ran F.A., Cong L., Yan W.X., Scott D.A., Gootenberg J.S., Kriz A.J., Zetsche B., Shalem O., Wu X., Makarova K.S., Koonin E., Sharp P.A, & Zhang F. (2015). In vivo genome editing using Staphylococcus aureus Cas9. Nature, 520(7546), 186-191.
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3

Quantification of Serum PCSK9 Levels

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Levels of serum PCSK9 were measured using the Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA kit (R&D Systems) according to manufacturer’s guidelines. Briefly, serum samples were diluted 1:200 in Calibrator diluent and allowed to bind for 2 h onto microplate wells that were precoated with the capture antibody. Samples were then sequentially incubated with PCSK9 conjugate followed by the PCSK9 substrate solution with extensive intermittent washes between each step. The amount of PCSK9 in serum was estimated colorimetrically using a standard microplate reader (BioRad iMark).
Moreno A.M., Palmer N., Alemán F., Chen G., Pla A., Jiang N., Chew W.L., Law M, & Mali P. (2019). immune-orthogonal orthologues of AAV capsids and of Cas9 circumvent the immune response to the administration of gene therapy. Nature biomedical engineering, 3(10), 806-816.
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4

Quantifying Plasma PCSK9 and LDL

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Blood samples (∼100 μL) were collected by retro-orbital bleeding 3 h after transfusion at day 0 and at time points indicated in the text for the sortagging approach, or for transplanted mice every month after bone marrow reconstitution. Plasma from these samples were isolated and stored at -80°C after quick freezing with liquid nitrogen until all samples were collected for simultaneous analysis. The plasma samples were diluted (20 μL in 200 μL PBS) then plasma PCSK9 levels quantified using a Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit (R&D systems) and plasma LDL levels measured using Cholesterol Assay Kit—HDL and LDL/VLDL (Abcam #ab65390). The detailed procedures were provided with the kit.
Deshycka R., Sudaryo V., Huang N.J., Xie Y., Smeding L.Y., Choi M.K., Ploegh H.L., Lodish H.F, & Pishesha N. (2021). Engineered red blood cells carrying PCSK9 inhibitors persistently lower LDL and prevent obesity. PLoS ONE, 16(11), e0259353.
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5

Evaluating PCSK9 and ANGPTL3 Levels in Mice

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Initial blood samples were collected following a 4 h fast. Age-matched littermates/colonymates were randomly assigned to experimental groups and administered AAV particles (n = 5) via retro-orbital injection. Blood samples were collected following a 4 h fast at 1-week intervals via the tail tip. After 4 weeks, all mice were killed by carbon dioxide asphyxiation after a 4 h fast. Whole livers were collected for genomic DNA isolation and analysis and for hematoxylin/eosin staining, and terminal blood samples were collected.
Pre-treatment and post-treatment plasma human PCSK9, mouse Pcsk9 or mouse ANGPTL3 were measured using the human proprotein convertase 9/PCSK9 Quantikine ELISA kit, mouse proprotein convertase 9/PCSK9 Quantikine ELISA kit or human angiopoietin-like 3 Quantikine ELISA kit, respectively, according to the manufacturer’s instructions (R&D Systems). Total cholesterol or triglyceride levels were measured using the Infinity cholesterol reagent or Infinity triglycerides reagent, respectively, according to the manufacturer’s instructions (Thermo Fisher).
Davis J.R., Wang X., Witte I.P., Huang T.P., Levy J.M., Raguram A., Banskota S., Seidah N.G., Musunuru K, & Liu D.R. (2022). Efficient in vivo base editing via single adeno-associated viruses with size-optimized genomes encoding compact adenine base editors. Nature Biomedical Engineering, 6(11), 1272-1283.
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6

Quantification of Serum PCSK9 Levels

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Levels of serum PCSK9 were measured using the Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA kit (R&D Systems) according to manufacturer’s guidelines. Briefly, serum samples were diluted 1:200 in Calibrator diluent and allowed to bind for 2 h onto microplate wells that were precoated with the capture antibody. Samples were then sequentially incubated with PCSK9 conjugate followed by the PCSK9 substrate solution with extensive intermittent washes between each step. The amount of PCSK9 in serum was estimated colorimetrically using a standard microplate reader (BioRad iMark).
Moreno A.M., Palmer N., Alemán F., Chen G., Pla A., Jiang N., Chew W.L., Law M, & Mali P. (2019). immune-orthogonal orthologues of AAV capsids and of Cas9 circumvent the immune response to the administration of gene therapy. Nature biomedical engineering, 3(10), 806-816.
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7

Multiplex Cytokine Profiling and Enzyme Analysis

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Plasma cytokines (IL-1β, IL-12p70, IFN-γ, IL-6, mKC, IL-10, and TNF-α) were measured with a Multi-Spot® 96-Well-7 Spot ELISA (Meso Scale Discovery, USA) [24 (link)]. Sandwich-type ELISA from eBioscience (USA) was used to measure TGF-β and MCP-1. Plasma PCSK9 was assessed with a Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA kit (R&D Systems, Inc., USA). COX activity in tissue extracts was measured with COX Fluorescent Activity Assay Kit (Cayman Chemicals, USA).
Sorokin A.V., Yang Z.H., Vaisman B.L., Thacker S., Yu Z.X., Sampson M., Serhan C.N, & Remaley A.T. (2016). Addition of Aspirin to a Fish Oil Rich Diet Decreases Inflammation and Atherosclerosis in ApoE-Null Mice. The Journal of nutritional biochemistry, 35, 58-65.
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8

AAV-mediated PCSK9 Modulation in Mice

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Male C57BL/6 (SLAC laboratory) mice at the age of 8 weeks were used for hydrodynamic tail vein injection. Mice were infected with 1.0×10 11 transducing units (TU) of AAV in 150 μL PBS by intravenous injection. Mice were sacrificed at 4 weeks post-injection of AAV. Before whole blood collection, mice were fasted for 4 hours. Then whole blood was collected at the time of euthanasia. Whole blood was stood at room temperature for 1 hour and centrifuged at 2000 g for 20 min. Then transfer the serum into new tubes for further analysis. Serum PCSK9 protein were measured with Mouse Proprotein Convertase9/PCSK9 Quantikine ELISA Kit (R&D Systems), according to the manufacturer's instructions. Serum parameters of liver functions, including alanine aminotransferase (ALT), aspartate transaminase (AST), were measured by automatic biochemical analyzer at the Adicon Clinical Laboratories.inc (Shanghai, China). Mouse primary hepatocytes were isolated by standard two-step collagenase perfusion and purified by 40% Percoll (Sigma) at low-speed centrifugation (1000 rpm, 10 min). Hepatocytes were resuspended in DMEM plus 10% fetal bovine serum (FBS) for FACS, and specific cell populations were used for RNA extraction.
Tong H., Huang J., Xiao Q., He B., Dong X., Liu Y., Yang X., Han D., Wang Z., Wang X., Ying W., Zhang R., Wei Y., Xu C., Zhou Y., Li Y., Cai M., Wang Q., Xue M., Li G., Fang K., Zhang H, & Yang H. (2023). High-fidelity Cas13 variants for targeted RNA degradation with minimal collateral effects. Nature biotechnology, 41(1).
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9

PCSK9 Quantification with ELISA

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PCSK9 protein level was assessed using the Mouse Proprotein Convertase 9/PCSK9 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN). Briefly, plasma was diluted and assayed by the ELISA sandwich technique. Plasma concentrations were estimated by calculation using a standard curve containing recombinant PCSK9.
Vergès B., Duvillard L., Pais de Barros J.P., Bouillet B., Baillot-Rudoni S., Rouland A., Petit J.M., Degrace P, & Demizieux L. (2021). Liraglutide Increases the Catabolism of Apolipoprotein B100-Containing Lipoproteins in Patients With Type 2 Diabetes and Reduces Proprotein Convertase Subtilisin/Kexin Type 9 Expression. Diabetes care, 44(4).
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