Journal: Journal of Lipid Research

Article Title: Development of a sensitive ELISA to quantify apolipoprotein CIII in nonhuman primate serum

doi: 10.1194/jlr.d011148

Figure Lengend Snippet: Fig. 1. ApoCIII ELISA can accurately detect and quantitate both human and nonhuman primate apoCIII. A-D: Purifi ed apoCIII from cynomolgus monkey and human serum were tested using both the ELISA de- scribed in this article and a commercially available apoCIII ELISA kit. Standard curves were generated to assess accurate detection of serum apoCIII. E, F: Western blots validate serial dilutions of purifi ed apoCIII protein tested by the ELISA methods.

Article Snippet: Selection of antibody pairs for ELISA Anti-human apoCIII antibodies were purchased from Novus and Abcam; details of the antibodies are listed in Table 1 .

Techniques: Enzyme-linked Immunosorbent Assay, Generated, Western Blot

Journal: Journal of Lipid Research

Article Title: Development of a sensitive ELISA to quantify apolipoprotein CIII in nonhuman primate serum

doi: 10.1194/jlr.d011148

Figure Lengend Snippet: Fig. 2. Determination of apoCIII ELISA sensitivity and linearity. A, B: ApoCIII ELISA was performed on serially diluted human se- rum and cynomolgus serum. In brief, we carried out a 4-fold serial dilution of serum samples in PBS containing 0.1% Tween-20, and then 50 m l/well of each dilution was tested by ELISA. We tested the ELISA sensitivity using cynomolgus serum samples serially diluted to represent decrements of 10% in apoCIII protein. C, D: Compari- son of ELISA and Western blot on a set of cynomolgus serum sam- ples that have 10% difference in apoCIII concentration.

Article Snippet: Selection of antibody pairs for ELISA Anti-human apoCIII antibodies were purchased from Novus and Abcam; details of the antibodies are listed in Table 1 .

Techniques: Enzyme-linked Immunosorbent Assay, Serial Dilution, Western Blot, Concentration Assay

Journal: Journal of Lipid Research

Article Title: Development of a sensitive ELISA to quantify apolipoprotein CIII in nonhuman primate serum

doi: 10.1194/jlr.d011148

Figure Lengend Snippet: Fig. 3. CP-900691, a PPAR- a agonist, decreases serum apoCIII protein. Type 2 diabetic monkeys were treated daily with 3 mg/kg CP-900691 for six weeks. Pretreatment and post-treatment sera were analyzed with the ELISA described in this article and Western blot. Results demonstrate that the ELISA can identify signifi cant de- creases in apoCIII protein, which was confi rmed by Western blot.

Article Snippet: Selection of antibody pairs for ELISA Anti-human apoCIII antibodies were purchased from Novus and Abcam; details of the antibodies are listed in Table 1 .

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Biology Open

Article Title: Apolipoprotein CIII regulates lipoprotein-associated phospholipase A 2 expression via the MAPK and NFκB pathways

doi: 10.1242/bio.201410900

Figure Lengend Snippet: (A,B) THP-1 cells were incubated with the indicated concentrations of apo CIII for 48 h. Lp-PLA 2 expression was tested by western blotting ( n = 3). (C,D) THP-1 cells were incubated with 30 µg apo CIII for the indicated durations. The proteins were then collected, and Lp-PLA 2 expression was examined by western blotting. (E) THP-1 cells were incubated with 30 µg apo CIII for the indicated durations. Lp-PLA 2 mRNA was subsequently extracted and detected using quantitative PCR ( n = 3). (F,G) THP-1 cells were transfected with a human apo CIII vector for 48 h ( n = 3). The Lp-PLA 2 activity was assayed using the PAF-AH assay kit, and the Lp-PLA 2 mRNA level was determined using quantitative PCR. * P <0.05, ** P <0.01, *** P <0.0001.

Article Snippet: The levels of proinflammatory cytokines which stimulated by apo CIII and released in the culture media were evaluated by Human IL-6 ELISA Kit, Human TNFα ELISA Kit and Human MCP-1 ELISA Kit (Boster Inc., China), following the manufacturer's instructions.

Techniques: Incubation, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Activity Assay

Journal: Biology Open

Article Title: Apolipoprotein CIII regulates lipoprotein-associated phospholipase A 2 expression via the MAPK and NFκB pathways

doi: 10.1242/bio.201410900

Figure Lengend Snippet: (A–C) THP-1 cells were incubated with 30 µg of apo CIII for the indicated durations. MCP-1, TNF-α and IL-6 were detected in the medium using an ELISA kit ( n = 3). (D–F) THP-1 cells were pretreated with 1-linoleoyl glycerol (75 µM) for 1 h and incubated with or without apo CIII (30 µg) for the indicated durations. The secretion of MCP-1, TNF-α and IL-6 was assayed using an ELISA kit ( n = 3). Error bars represent mean±s.e.m. *** P <0.0001.

Article Snippet: The levels of proinflammatory cytokines which stimulated by apo CIII and released in the culture media were evaluated by Human IL-6 ELISA Kit, Human TNFα ELISA Kit and Human MCP-1 ELISA Kit (Boster Inc., China), following the manufacturer's instructions.

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Biology Open

Article Title: Apolipoprotein CIII regulates lipoprotein-associated phospholipase A 2 expression via the MAPK and NFκB pathways

doi: 10.1242/bio.201410900

Figure Lengend Snippet: THP-1 cells were incubated with apo CIII and the indicated inhibitors. (A–C) After incubating with apo CIII for the indicated durations, THP-1 cell proteins were collected to detect p42/44 and p65 expression and phosphorylation using western blotting ( n = 3). (D–G) THP-1 cells were incubated in PD98059 and PDTC for 1 h and then incubated with apo CIII, PD98059 and PDTC for an additional 12 h. The levels of Lp-PLA 2 , p42/44, p65 and their phosphorylated counterparts were determined by western blotting ( n = 3). (H) THP-1 cells were treated as described in D, except that the incubation time was extended to 36 h. Quantitative PCR was performed to determine the levels of Lp-PLA 2 mRNA expression ( n = 3). Error bars represent mean±s.e.m. * P <0.05, ** P <0.01.

Article Snippet: The levels of proinflammatory cytokines which stimulated by apo CIII and released in the culture media were evaluated by Human IL-6 ELISA Kit, Human TNFα ELISA Kit and Human MCP-1 ELISA Kit (Boster Inc., China), following the manufacturer's instructions.

Techniques: Incubation, Expressing, Phospho-proteomics, Western Blot, Real-time Polymerase Chain Reaction

Journal: Biology Open

Article Title: Apolipoprotein CIII regulates lipoprotein-associated phospholipase A 2 expression via the MAPK and NFκB pathways

doi: 10.1242/bio.201410900

Figure Lengend Snippet: Blood samples from apo CIII transgenic and wild-type pigs were collected, and the plasma was isolated for use in Lp-PLA 2 activity assays. Mononuclear cells were isolated for total RNA extraction. (A) The plasma Lp-PLA 2 activity in the wild-type and apo CIII transgenic pigs was assayed less than 2 h after plasma isolation. (B) Total RNA was extracted from the mononuclear cells, and quantitative PCR for Lp-PLA 2 mRNA level was conducted. (C) The wild-type and transgenic pigs were fasted for 16 h and fed olive oil for 2 h, the plasma was isolated, and the Lp-PLA 2 activity were measured. n = 8 in the wild-type pigs, and n = 4 in the apo CIII transgenic pigs. Error bars represent mean±s.e.m. (D) Lp-PLA 2 stable expressed HepG2 cells was incubated with 1-linoleoyl glycerol for 12 h. Quantitative PCR was performed to determine the levels of apo CIII mRNA expression ( n = 3). (E) Blood and liver tissue were collected from slaughtered landraces, and quantitative PCR was performed to determine the levels of apo CIII mRNA expression in liver tissue and Lp-PLA 2 mRNA expression in monocytes isolated from blood. Linear regression analysis is shown. * P <0.05, ** P <0.01, *** P <0.0001. (F) A schematic model of apolipoprotein CIII regulates Lp-PLA 2 expression.

Article Snippet: The levels of proinflammatory cytokines which stimulated by apo CIII and released in the culture media were evaluated by Human IL-6 ELISA Kit, Human TNFα ELISA Kit and Human MCP-1 ELISA Kit (Boster Inc., China), following the manufacturer's instructions.

Techniques: Transgenic Assay, Clinical Proteomics, Isolation, Activity Assay, RNA Extraction, Real-time Polymerase Chain Reaction, Incubation, Expressing

Journal: Inhalation Toxicology

Article Title: Study of cardiovascular disease biomarkers among tobacco consumers, part 2: biomarkers of biological effect

doi: 10.3109/08958378.2015.1013227

Figure Lengend Snippet: Serum lipid biomarkers.

Article Snippet: Total cholesterol, triglycerides, low-density lipoprotein cholesterol (LDL-C), HDL-cholesterol (HDL-C), very LDL-cholesterol (VLDL-C), apolipoprotein A1 (Apo A1), apolipoprotein A2 (Apo A2), apolipoprotein B100 (Apo B100), apolipoprotein(a) [Lp(a)], oxidized LDL (ox-LDL), folate, fibrinogen and tissue inhibitor of matrix metalloproteinase-1 (TIMP1) were measured at Pacific Biomarkers Inc. (Seattle, Washington). α 1 -Antitrypsin (AAT) was measured at ARUP Laboratories (Salt Lake City, Utah).

Techniques: Biomarker Discovery, Comparison