The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use a concentration of 10 - 20 µg/ml.
Use at an assay dependent concentration.
Use a concentration of 0.5 - 4 µg/ml. Predicted molecular weight: 24 kDa.Can be blocked with GDNF peptide (ab51926).
Western blot may give multiple bands. GDNF homodimerizes and has 5 isoforms along with a signal and propeptide.
Use a concentration of 10 - 20 µg/ml.
Neurotrophic factor that enhances survival and morphological differentiation of dopaminergic neurons and increases their high-affinity dopamine uptake.
In the brain, predominantly expressed in the striatum with highest levels in the caudate and lowest in the putamen.
Involvement in disease
Defects in GDNF may be a cause of Hirschsprung disease (HSCR) [MIM:142623]. In association with mutations of RET gene, defects in GDNF may be involved in Hirschsprung disease. This genetic disorder of neural crest development is characterized by the absence of intramural ganglion cells in the hindgut, often resulting in intestinal obstruction. Defects in GDNF are a cause of congenital central hypoventilation syndrome (CCHS) [MIM:209880]; also known as congenital failure of autonomic control or Ondine curse. CCHS is a rare disorder characterized by abnormal control of respiration in the absence of neuromuscular or lung disease, or an identifiable brain stem lesion. A deficiency in autonomic control of respiration results in inadequate or negligible ventilatory and arousal responses to hypercapnia and hypoxemia.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GDNF antibody (ab18956)This image is courtesy of an Abreview submitted by Chi-Li Lin.
ab18956 staining GDNF in rat brain tissue sections by IHC-P (Formalin/PFA-fixed paraffin-embedded sections). Cells were fixed with paraformaledhyde, permeabilized with 0.1% Tween-20 and PBS and blocked with 1% BSA for 30 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer pH 6.0. Samples were incubated with primary antibody (1/150 in TBS) for 24 hours at 4oC. ab96883 at a dilution of 1/200 was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-GDNF antibody (ab18956)This image is courtesy of an abreview submitted by Tamas Bellak, University of Szeged.
Immunocytochemistry/ Immunofluorescence analysis of genetically modified rat embryonic fibroblasts labeling GDNF with ab18956 at 1/150 dilution. Cells were fixed in formaldehyde and permeabilized with Tx-100. Staining with ab18956 at 1/150 was carried out for 14 hours at 4°C in PBS buffer. A Goat Anti-Rabbit (Alexa Fluor® 594) secondary antibody was used at 1/600 dilution.
ICC/IF image of ab18956 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18956, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of ab18956 staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18956, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.