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Notes: These authors investigated the mechanism of action of the migraine mediator calcitonin gene-related peptide (CGRP), which is known to sensitize the P2X₃ pain receptors to increase impulse flow to brain stem trigeminal nuclei. They showed that CAM KII inhibitors prevented CGRP-induced upregulation of P2X₃ mRNA using real-time RT-PCR, and then confirmed that active CAM KII was involved in the signaling mechanism by staining with Anti-ACTIVE^® CAM KII Antibodies. The immunoreactivity was upregulated by CGRP treatment of trigeminal neurons, and the distribution of the active CAM KII was localized to the membrane region. They then examined the mechanism of transcriptional activation of the P2X₃ pain receptor genes and showed that the transcription factor CREB, which is known to be dependent on CAM KII for activation and nuclear localization, was involved. The authors also investigated the role of BDNF in the process, because CGRP is known to promote BDNF expression by trigeminal neurons, and because BDNF is thought to be involved in pain processing. They used the BDNF E_max Immunoassay System to measure BDNF levels in culture media after application of CGRP to neuronal cell cultures, and demonstrated that there was a fourfold increase in BDNF after CGRP exposure. They also showed that BDNF promoted CAMKII and CREB activation. (3873)

Notes: This study showed that the heat-shock DnaJ-containing protein 1b (HSJ1b) and cystamine are neuroprotective in Huntington disease and act by increasing release of brain-derived neurotrophic factor (BDNF), which is critical for the survival of striatal neurons. Mouse neuronal cells were transfected with vectors expressing BDNF, HSJ1b, or with a negative control (empty) vector. HSJ1b was shown to significantly increase BDNF release. Treatment of cells with cystamine, a candidate drug for treatment of Huntington disease, also increased BDNF release. RNA interference experiments showed that in cells where HSJ1b expression was reduced, cystamine no longer stimulated BDNF release, indicating that HSJ1b was required for the protective effect mediated by cystamine. In all these experiments the amount of BDNF in the cultured cell supernatants was measured using the BDNF E_max^® ImmunoAssay System. (3480)

Notes: In this study, the BDNF E_max^® ImmunoAssay System was used to measure BDNF levels in heparinized bone marrow plasma from patients with multiple myeloma (MM) and in control samples. The ELISA analysis was used to establish that BDNF is secreted from primary MM cells in vivo. (3474)

Notes: This study investigated the angiogenic potential of brain-derived neurotrophic factor (BDNF) in ischemic and non-ischemic tissues of adult mice. As part of this study, recombinant Adenovirus encoding rat BDNF was administered to mice via tail-vein injection, and BDNF levels in plasma were then monitored using the BDNF E_max^® ImmunoAssay System. (3473)

Notes: This paper describes an assay to test BDNF dissociation from FluoSpheres (amine-modified microspheres, 1μM diameter; Molecular Probes).  BDNF was covalently attached to the microspheres.  Release of BDNF was tested in supernatants from the BDNF-microspheres in PBS,  in complete media, and in culture with rat Dorsal root ganglia.  Promega’s BDNF Emax^® ImmunoAssay System was used to determine the amounts of BDNF in these samples.  The DeadEnd™ Fluorometric TUNEL System was used to examine rat Dorsal root ganglia cells that had been transfected with either dynamitin or β-Galactosidase.  In the TUNEL experiments, cells were fixed in 4% paraformaldehyde and counterstained with DAPI.  Cells were also stained with Promega’s Anti-β-Galactosidase, Purified Monoclonal Antibody.  For these immunocytochemical stains, a 1:500 dilution of Anti-β-Galactosidase antibody and a 1:1,500 dilution of a secondary antibody conjugated to Alexa Fluor-488 (Molecular Probes) were used.  The stains were visualized by fluorescence microscopy.  (3055)

Notes: In this study of factors that may contribute to Familial Alzheimer's disease, transgenic mice expressing mutant amyloid precursor protein and mutant presenilin 1 protein were constructed. The levels and effects of several neurotransmitters and BDNF were evaluated by either HPLC analysis or by immunodetection using the BDNF E_max® ImmunoAssay System. The authors describe a method for extraction of BDNF from mouse brain that gives a recovery rate of approximately 70% compared to 2-5% recovery with the standard procedure as given in the BDNF E_max® ImmunoAssay System protocol.  (3121)

Notes: The authors examine the role of brain-derived neurotrophic factor (BDNF) in alcohol addition. RNA was isolated from primary rat hippocampal neurons cultured in the absence or presence of ethanol. The RNA was reverse transcribed using the Reverse Transcription System, and BDNF and GPDH RNAs were quantitated by fluorescent real-time PCR. BDNF and nerve growth factor (NGF) protein levels were monitored using the BNDF and NGF Emax^® ImmunoAssay Systems. (3441)

Notes: To study the role of bone morphogenetic protein (BMP) in the development of peripheral sensory neurons, transgenic mice were created that overexpressed either a BMP inhibitor, noggin, or a ligand, BMP4. To determine if the increased trigeminal innervation and neuron numbers seen in these mice were due to increased neurotrophin levels, the mystacial (whisker) pads of 3-day-old mice were tested for the presence of NGF, BDNF and NT-3 using each of the corresponding E_max^® ImmunoAssay Systems. The results indicated there were no significant changes in neurotrophin levels except for a decrease in NT-3 in the noggin transgenic mice. (3337)

Notes: Adult male rhesus monkeys were divided into two groups to model the effect of calorie-restriction on the progress of Parkinson’s disease. Caudate nucleus samples from both sides of the brain were taken 16–18 weeks after inducing hemiparkinsonism. Levels of neurotrophins were measured using the BDNF and GDNF E_max^® ImmunoAssay Systems. The amount of BDNF and GDNF were compared to the total protein present in each sample and expressed as picogram per milligram units. (3335)

Notes: Cultured hippocampal slices were prepared from Sprague-Dawley rat pups (9–10 days postnatal). After exposure to positive AMPA receptor modulators, the slices were homogenized and total BDNF protein content measured using the BDNF Emax^® ImmunoAssay System. (2812)

Notes: BDNF levels in mouse brains were evaluated using the BDNF Emax^® ImmunoAssay System and compared with with BDNF levels in spinal cords of end-stage ALS.  Samples were not acid treated to allow  measurement of free, mature BDNF. (2808)

Notes: Both the BDNF Emax^® ImmunoAssay System and the NT-3 Emax^® ImmunoAssay System were used to determine neurotrophin levels in olfactory bulb nerve layer isolated from adult female Fischer F344 rats. The primary cells were infected with adenovirus vectors expressing BDNF and NT-3 and the media assayed 3 days later. (2803)

Notes: In this paper, the BDNF Emax^® ImmunoAssay System was used to analyze BDNF levels in homogenized retina tissue from New Zealand White Rabbits. (2817)

Notes: The BDNF Emax^® ImmunoAssay System and the NGF Emax^® ImmunoAssay System were used to assay the levels of neurotrophinin in male Long–Evans rat brains. After cocaine consumption, brain tissue was homogenized and the BDNF and NGF levels assayed. (2805)

Notes: The BDNF Emax^® ImmunoAssay System was used with human follicular fluid, granulosa cell culture media oocyte, or embryo culture to measure the amount of BDNF. (2801)

Notes: The BDNF Emax^® ImmunoAssay System was used with dorsal horn tissue derived from lumbar spinal cord of adult rats. The amount of BDNF was measured after electrical stimulation using low and high frequency pulses. (2800)

Notes: Transplantation of olfactory ensheathing cells into lesions in the central nervous system is able to stimulate the growth of axons and in some cases restore functional connections. To investigate the mechanism, the authors quantitated the production of growth factors and expression of corresponding receptors by rat olfactory ensheathing cells. Levels of brain-derived neurotrophic factor, glial-cell-derived neurotrophic factor, nerve growth factor, and neurotrophin-3 were monitored using Promega's BDNF E_max® ImmunoAssay System, GDNF E_max® ImmunoAssay System, NGF E_max® ImmunoAssay System, and NT-3 E_max® ImmunoAssay System, respectively.  (2318)

Notes: Brain-derived neurotrophic factor, glial cell line-derived neurotrophic factor, and neurotrophin-3 protein levels were quantitated in the striatum and ventral midbrain of young and aged rats following a lesion of the nigrostriatal system. BDNF, GDNF, and neurotrophin-3 levels were monitored using Promega's BDNF E_max® ImmunoAssay System, GDNF E_max® ImmunoAssay System and NT-3 E_max® ImmunoAssay System, respectively. (2324)

Notes: Regional and temporal patterns of neurotrophin messenger RNA and protein levels were characterized in the developing rat hippocampus, neocortex and cerebellum on postnatal days 1, 7, 14, 21, and 92. Brain-derived neurotrophic factor nerve growth factor, and neurotropin-3 protein levels were determined using Promega's BDNF E_max® ImmunoAssay System, NGF E_max® ImmunoAssay System, and NT-3 E_max® ImmunoAssay System. (2319)

Notes: The effect of neuromuscular activity on neurotrophin expression in the lumbar spinal cord region and innervating skeletal muscle of adult rats was examined. BDNF protein levels in the soleus muscle after exercise were determined using Promega's BDNF E_max® ImmunoAssay System. (2321)

Notes: Used NT-3 Emax^® ImmunoAssay System, NT-4 Emax^® ImmunoAssay System, BDNF Emax^® ImmunoAssay System and NGF Emax^® ImmunoAssay System to monitor secreted neurotrophins from human lamina cribrosa cells and human optic nerve head astrocytes.  After a 72-hour treatment with serum-free medium containing 0.5 mg/ml BSA, the media was removed and then assayed for the respective neurotrophin. (2796)

Notes: The regulation of brain-derived neurotrophic factor, nerve growth factor and other neurotrophin ligands and receptors by estrogen was characterized in young adult and reproductively senescent rat brains. BNDF, NT-4, and NGF protein levels were monitored using Promega's BDNF E_max® ImmunoAssay, NT-4 E_max® ImmunoAssay System, and NGF E_max® ImmunoAssay Systems, respectively . (2320)

Notes: Sprague-Dawley rat neuronal cells were cultured and exposed to hypoxia. The neuronal cultures were fractionated into cytosol, supernatant and ER-rich components, and the amount of BDNF determined using the BDNF Emax^® ImmunoAssay System. (2811)

Notes: A rat model of Huntington's disease was employed to determine whether neurotrophins are able to promote the survival of striatal projection neurons in vivo. Results suggest that BDNF might be beneficial for the treatment of striatonigral degenerative disorders, including Huntington's disease. BDNF and NT-3 levels in rat striatum were quantitated using Promega's BDNF E_max® ImmunoAssay System and NT-3 E_max® ImmunoAssay System, respectively. (2323)

Notes: Ethanol has been previously shown to inhibit the anti-apoptotic effect of NMDA.  The decreased anti-apoptotic effect of NMDA was associated with a number of factors including decreased induction of brain-derived neurotrophic factor expression. Levels of BDNF in cultured rat cerebral granular neurons were quantitated using Promega's BDNF E_max® ImmunoAssay System. (2316)