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Apelin 12

Manufactured by Phoenix Pharmaceuticals
Sourced in United States

Apelin-12 is a laboratory research reagent used for scientific experiments. It is a 12-amino acid peptide that functions as a ligand for the apelin receptor, which is involved in various physiological processes. The core function of Apelin-12 is to serve as a tool for researchers to study the role and mechanisms of the apelin receptor and its signaling pathways in different biological systems.

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5 protocols using apelin 12

1

Biomarker Analysis in Blood Samples

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Blood samples were collected into EDTA and serum tubes and were centrifuged for 15 min at 1000g at 4°C and 21°C, respectively. Aliquots were immediately frozen in liquid nitrogen and subsequently stored at −80°C until analysis. Plasma glucose, FFA, TAG and total cholesterol were analyzed with standard enzymatic methods (ABX Pentra 400 autoanalyzer, Horiba ABX, Montpellier, France). Plasma glycerol was analyzed with an enzymatic assay (Enzytec Glycerol, Roche Biopharm, Basel, Switzerland) automated on a Cobas Fara spectrophotometric autoanalyzer. Plasma insulin, leptin, and adiponectin concentrations were analyzed with commercially available radioimmunoassay (RIA) kits (Human insulin specific RIA, human leptin RIA, human adiponectin RIA, Millipore Corporation, Billerica, MA). Plasma concentrations of IL‐6 (Meso Scale Discovery, Gaithersburg, MD), apelin‐12 (Phoenix pharmaceuticals Inc., Burlingame, CA), RBP4 (R&D systems, Minneapolis, MN) and Vaspin (Adipogen, San Diego, IL) were determined by ELISA. The apelin‐12 ELISA has 100% cross‐reactivity with human apelin‐12, apelin‐13, and apelin‐36. Serum ACE activity was measured with a standard enzymatic assay (Bühlmann, Basel, Switzerland) while serum nesfatin‐1 (Phoenix pharmaceuticals Inc., Burlingame, CA) concentrations were measured by ELISA.
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2

Cytokine Profiling in Cell Culture

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Supernatants of cell culture medium were collected after the experiment. According to the protocol of the manufacturer, expression of Apelin-12 (phoenix pharmaceuticals, USA), tumor necrosis factor-α (TNF-α), interleukin-1beta (IL-1β) and interleukin-6 (IL-6) (Boster, Wuhan, China) were detected in the supernatant.
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3

Serum Apelin Concentration Assessment

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The peripheral blood samples were collected from all participants after 12-h overnight fast. The serum Apelin concentration was measured by an ELISA kit (Apelin-12, Phoenix pharmaceuticals, Belmont, USA) according to manufacturer’ protocol. The sensitivity was 0.05 ng/mL, and intra- and inter-assay variations were <5 and <10 %, respectively.
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4

Apelin and Inflammatory Biomarkers

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The peripheral blood samples were collected from all participants after 12-h overnight fast. Apelin was measured by an ELISA kit (Apelin-12, Phoenix pharmaceuticals, Belmont, USA). The sensitivity was 0.05 ng/mL, and intra- and inter-assay variations were <5 and <14 %, respectively. Serum high-sensitivity C-reactive protein (hs-CRP) measurements were performed using a commercially available high-sensitivity assay (Roche Diagnostics).
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5

Apelin Receptor Characterization in CHO and HUVEC Cells

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Chinese hamster ovary cells and human umbilical vein endothelial cells were obtained from American Type Culture Collection (ATCC). All restriction enzymes were from NewEngland BioLabs. [Ala13] apelin-13, apelin-12, 13, 15, 17, 36 were purchased from Phoenix Pharmaceuticals. Lipofectamine 2000, streptavidin-phycoerythrin (SA-PE) and Opti-MEM I were obtained from Invitrogen Life Technologies. Isopropyl-beta-D-thiogalactopyranoside (IPTG) and Coelenterazine h were obtained from Promega. HEPES-buffered phenol red−free medium, Dulbecco’s modified eagle medium and Incomplete RPMI-1640 culture medium were purchased from Gibco. Anti-HA-agarose was obtained from Pierce Chemical Co. Polyclonal horseradish peroxidase-conjugated goat anti-rabbit immuno-globulins/HRP was obtained from Zhong Shan Gold Bridge Biology Corporation (China). Anti-Myc antibody and anti-HA antibody were purchased from Cell Signaling Technology
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