JCI - PAR-1 contributes to the innate immune response during viral infection
Life Diagnostics). Plasma TAT levels were quantified by ELISA (TAT Enzygnost Micro Kit; Dade Behring/Siemens). ... Measurement of plasma cardiac troponin I and TAT complexes. Blood was collected from the inferior vena cava into the ... The Vecastatin ABC Kit (Vector Laboratories) was used to detect the biotin-tagged secondary antibody (14). Images were taken ... In vitro experiments with CFs showed that activation of PAR-1 enhanced poly I:C activation of p38 and expression of IFN-β and ...
JCI Insight - Ginger intake suppresses neutrophil extracellular trap formation in autoimmune mice and healthy humans
Neutrophils, peripheral blood mononuclear cells (PBMCs), and plasma were isolated. Compared with pre-ginger samples, there was ... The activity of PDE was measured using the Bridge-It cAMP-PDE assay kit (Mediomics, catalog PD-1016). The supernatant was mixed ... This was followed by the ImmPRESS HRP Goat Anti-Rat IgG Polymer Detection Kit (Vector Laboratories) with diaminobenzidine as ... This protocol used several reagents from the Cell Death Detection ELISA kit (Roche). First, a high-binding EIA/RIA 96-well ...
JCI - PTG gene deletion causes impaired glycogen synthesis and developmental insulin resistance
Upon digestion, protein concentration of an aliquot was determined by Bradford assay (Bio-Rad Laboratories Inc.). The sample ... Mixed arterial/venous blood was collected from the tail tip at 0, 15, 30, 60, 90, and 120 minutes. Blood glucose concentration ... Total RNA (5 μg) was then used for RT-PCR using the cDNA Cycle kit from Invitrogen Corp. The primers used for cDNA synthesis ... The specific activity of glucose in serum was determined as follows: a plasma sample was deproteinized with 67 volumes of 3.5% ...
JCI Insight - Ginger intake suppresses neutrophil extracellular trap formation in autoimmune mice and healthy humans
Neutrophils, peripheral blood mononuclear cells (PBMCs), and plasma were isolated. Compared with pre-ginger samples, there was ... The activity of PDE was measured using the Bridge-It cAMP-PDE assay kit (Mediomics, catalog PD-1016). The supernatant was mixed ... This was followed by the ImmPRESS HRP Goat Anti-Rat IgG Polymer Detection Kit (Vector Laboratories) with diaminobenzidine as ... This protocol used several reagents from the Cell Death Detection ELISA kit (Roche). First, a high-binding EIA/RIA 96-well ...
JCI - PPARγ regulates adipocyte cholesterol metabolism via oxidized LDL receptor 1
Measurement of serum oxLDL levels. Assays were performed using the Oxidized LDL ELISA immunoassay kit as described by the ... Regulation of fasted blood glucose by resistin. Science. 2004. 303:1195-1198. View this article via: PubMed Google Scholar ... PPARγ is required for the differentiation of adipose tissue in vivo and in vitro. Mol. Cell. 1999. 4:611-617. View this article ... In vivo experiments. C57BL/6J and ob/ob mice (Jackson Laboratory), aged 11 weeks, were housed (n = 4 per cage) under 12-hour ...
JCI - Desmosomal COP9 regulates proteome degradation in arrhythmogenic right ventricular dysplasia/cardiomyopathy
The first-strand cDNA was generated using PrimeScript RT Reagent Kit with gDNA Eraser (Takara). Real-time PCR was performed on ... In vitro reconstitution assays refining the minimal CSN subunits required for deneddylation activity defined the CSN5-CSN6 ... A new diagnostic test for arrhythmogenic right ventricular cardiomyopathy. N Engl J Med. 2009;360(11):1075-1084.. View this ... Genetic testing on peripheral blood DNA was performed using a panel consisting of 7 known ARVD/C genes (PKP2, DSP, DSG2, DSC2, ...
JCI Insight - Defective jagged-1 signaling affects GnRH development and contributes to congenital hypogonadotropic hypogonadism
Protein content was assayed by the BCA protein assay kit (Thermo Fisher Scientific, 23225). Total proteins were fractionated by ... 10 mL Automated Blood kit (Invitrogen). The CHH gene panel was designed using Illumina Design Studio and included the following ... and propagated in vitro from human fetal OE (27). These cells have been shown to preserve cell motility in vitro (57). ... Antibody-protein complexes were then detected using the Novex ECL Chemiluminescent substrate reagent kit (Thermo Fisher ...
JCI Insight - Controlled human malaria infection with a clone of Plasmodium vivax with high-quality genome assembly
Cryopreservation and in vitro testing of P. vivax-infected blood. After blood donation, the leukodepleted blood from both ... C and transported immediately to the Jenner Institute Laboratories. RBC were separated from plasma by centrifugation (830g for ... vivax blood-stage parasitemia in volunteers blood in real time. The assay targets the 18S ribosomal RNA (rRNA) gene and was ... Following admission, a 250 mL blood sample was collected using aseptic technique, via a whole blood donation kit containing an ...