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Notes: To examine the effect of GDNF expression on body weight, recombinant adeno-associated virus (rAAV) expressing GDNF was injected intrahypothalamically into young and senescent rats. Hypothalamuses were harvested, lysed and used undiluted, and at dilutions of 1:50 and 1:500 in the GDNF E_max^® ImmunoAssay System. The amount of GDNF was expressed as a concentration of GDNF protein per milligram of hypothalamic tissue sample. (3342)

Notes: In this study, the GDNF E_max^® ImmunoAssay System was used to determine the GDNF content of various tissues used to treat Parkinson disease in animals and humans. Tissues tested included rodent carotid body, superior cervical ganglia, adrenal medulla and Zuckerkandl's organs. Tissues were frozen in liquid nitrogen, then homogenized. The ELISA was then performed according to the supplied protocol, with the following exceptions: the anti-GDNF mAb was used at a 1:500 dilution, and the anti-GDNF pAb was used at a 1:250 dilution. GDNF secreted from dissociated cells of the various tissue types was also evaluated in an in situ ELISA, where dissociated cells were cultured for 48 hours in plates coated with anti-GDNF mAb. The cells were then removed by washing and the amount of released GDNF evaluated by ELISA. (3475)

Notes: The GDNF Emax^® ImmunoAssay System was used to measure GDNF levels in adenovirus-infected rats.  The researchers describe analyzing GDNF levels in brain lysates from naïve, immunized and nonimmunized rats. Naïve animals displayed GDNF expression after infection with an adenovirus vector that expressed GDNF (rAAV2-GDNF). Data aredisplayed graphically as picograms of GDNF per milligram of brain tissue.  (3254)

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: Male mice were bilaterally injected with neural tracer cholera toxin subunit B into gastrocnemius muscles to selectively label motoneurons that retained axons. One week later, tissue was harvested and homogenized and theGDNF Emax^® ImmunoAssay System was used on acidified supernatants to measure GDNF levels. (2802)

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: In Alzheimer's disease, cholinergic neurons are lost from the medial septum and nucleus basalis of Meynert. Nerve growth factor and glial cell line-derived neurotrophic factor were examined to determine if these growth factors can rescue the cholinergic neurons. GDNF and NGF levels in the sections of the basal nucleus of Meynert in rat was determined using Promega's GDNF E_max® ImmunoAssay System and NGF E_max® ImmunoAssay System, respectively. Glial cell line-derived neurotrophic factor was capable of partially restoring the number of cholinergic neurons. (2325)

Notes: To determine the protective effects of glial cell line-derived neurotrophic factor on axotomized facial motoneurons, GDNF was continuously expressed within the rat facial nucleus and these rats were subjected to facial nerve lesion. Axotomized motoneurons were completely protected against cell death. Levels of GDNF in the brainstem were determined using Promega's GDNF E_max® ImmunoAssays System.  (2326)

Notes: Purified embryonic rat motoneurons were >95% transduced with HIV1-based lentiviral vectors encoding beta-galactosidase and human glial cell line-derived neurotrophic factor. GDNF levels in conditioned cell culture medium were determined using Promega's GDNF E_max® ImmunoAssay System. (2328)

Notes: Wildtype and mutant rat GDNF was expressed in African green monkey kidney cells (COS cells) and purified from the cell media. Quantities of the wildtype and mutant factors before and after purification were determined using the GDNF E_max^® ImmunoAssay System. (1456)

Notes: The Anti-Human GDNF pAb was used to detect GDNF in rat pituitaries by immunohistochemistry. To stain for GDNF alone, the rats were perfused with cold 4% paraformaldehyde, paraffin embedded, then sliced into 8µm sections. The sections were permeabilized with a brief trypsin treatment. The sections were blocked with normal donkey serum and incubated with the Anti-Human GDNF pAb (1:250 dilution) either overnight at ambient room temperature or for 48 hours at 4°C followed by colorimetric detection. For co-localization with other marker antibodies, the pituitaries were flash frozen and 10µm sections obtained. The frozen sections were fixed in 4% paraformaldehyde and permeablized with 0.3% Triton^® X-100 in 5% normal goat serum. The sections were treated for 48 hours at 4°C in the presence of the Anti-Human GDNF pAb (1:250 dilution) in the goat serum/0.3% Triton^® X-100 solution with accompanying antibodies. Actual levels of GDNF in the pituitaries were determined with the GDNF E_max^® ImmunoAssay System. (0053)

Notes: An adeno-associated viral vector was used for long-term expression of GDNF in either the nigral DA neurons, in the striatal target cells, or in both of these structures. Only rats expressing GDNF in the striatum displayed behavioral recovery, and significant reinnervation of the lesioned striatum. GDNF expression was monitored using Promega's GDNF E_max® ImmunoAssay System. (2329)

Notes: A lentiviral vector was used to express glial cell line-derived neurotrophic factor in nonhuman primate models of Parkinson's disease. GDNF expression in the substantia nigra seemed to reverse functional deficits and prevent nigrostriatal degeneration. GDNF expression in transduced and non-transduced cells of the rhesus monkey substantia nigra was quantitated using Promega's GDNF E_max® ImmunoAssay System. (2330)

Notes: In a Parkinson's disease model, the combination of adenoviral gene transfer of X-chromosome-linked inhibitor of apoptosis and of the glial cell line-derived neurotrophic factor to the rat striatum showed promise in preventing neuronal cell death and preserving neuronal function. GDNF levels in the striatum and in the human neuroblastoma cell line SH-Sy5Y were quantitated using Promega'sGDNF Emax® ImmunoAssay System.  (2327)

Notes: The GDNF Emax® ImmunoAssay System (GDNF ELISA) was used to determine the amount of GDNF secreted by 293T cells infected with a GDNF-encoding lentivirus. The Self-inactivating (SIN) lentiviral vector produced approximately 900ng of GDNF per day and the first generation lentiviral vector produced only about 150ng of GDNF per day. The SIN virus was injected into rat substantia nigra and punches of the mesencephalon including the substantia nigra were collected and analyzed for GDNF levels. The SIN virus produced about 2.5ng of GDNF per punch one week after injection and control animal given a SIN-lacZ construct had undetectable levels of GDNF. (1268)

Notes: The GDNF E_max^® ImmunoAssay System (GDNF ELISA) was used to investigate GDNF levels in rat cerebralspinal fluid after birth out to three weeks. The CSF levels of GDNF ranged from 71pg/ml to a high of 138pg/ml. The researchers used only samples with no contaminating blood. (0992)

Notes: The study makes extensive use of the GDNF E_max^® ImmunoAssay System to monitor the level of GDNF secreted into the media of rat C6 glioblastoma cells. The researchers validated that the system was good for GDNF levels between 10pg/ml and 500pg/ml. C6 cells without external stimuli secreted GDNF at an average of 36 ± 5.6 pg/ml (n = 66) and ranged from undetectable levels to 150pg/ml. The NO generator sodium nitroprusside increased GDNF secretion at least 5-fold. (GDNF ELISA) (0222)

Notes: The levels of GDNF, NGF, and NT-3 were quantitated in various mouse tissues, including spleen, muscle, heart, lung, liver, kidney, bladder, gastrointestinal tract, brain, and serum. Additional samples screened for NGF included pig, goat, horse, chicken, rabbit, sheep, and bovine serum and rat brain. Samples were untreated or acid treated by acidification to pH <3.0 followed by neutralization to pH 7.6. In all tissue samples and most serum samples, the detectable levels of GDNF, NGF and NT-3 increased up to 35-fold. It is thought that the acidification process promotes dissociation of receptors and associated proteins making the neurotrophins more accessible. Levels of GDNF, NGF, and NT-3 were determined using the GDNF, NGF, and NT-3 E_max ImmunoAssay Systems, respectively.  Please note that the coat antibody in the NGF E_max ImmunoAssay System was changed 12/2002 and may affect cross-reactivity. (3105)

Notes: BHK cells stably expressing rat GDNF were encapsulated in a polymer for implantation into the medial forebrain bundle and substantia nigra. The GDNF E_max^® ImmunoAssay System (GDNF ELISA) was used to determine the release rate of the encapsulated cells. A cell capsule was placed in 500µl of culture medium for 60 minutes and the supernatant assayed for GDNF. The capsule was found to release ~5ng GDNF/day. (0238)

Notes: The GDNF E_max^® ImmunoAssay System was used to measure GDNF levels in culture. GDNF was assayed from rat mesencephalic cells and astrocytes both pre- and post-infection with a recombinant adenovirus that produced GDNF. The same assay was also performed with HeLa cells. (1451)

Notes: The GDNF E_max ImmunoAssay System was used to quantify the level of GDNF in the conditioned media of HeLa cells transduced with a recombinant adenovirus expressing GDNF. The RQ1 DNase was used to treat RNA sample to remove genomic DNA and the AMV reverse transcriptase was used for first strand cDNA synthesis. (1605)