Overview
For long-term storage in solution, we recommend preparing a stock solution by dissolving the product in sterile water at a concentration of at least 0.1 mg/mL. Divide the stock solution into small aliquots and store at -20°C. Before use, thaw the relevant vial(s) and dilute to the desired working concentration in your working buffer. It is recommended to prepare fresh solutions in working buffers just before use. Repeat freeze-thawing may result in loss of activity.
- Live cell imaging of recombinant human GDNF-Biotin protein in live intact human SH-SY5Y cells.(A) CellMask™ Actin 1X solution was applied for 30 minutes at 370C, resulting in a green fluorescence to visualize cellular membrane. (B) Following this, the same cells underwent incubation with 0.1 µM of recombinant human GDNF protein-Biotin (#G-240-B) for 30 minutes at 370C, followed by Alexa Streptavidin-647, leading to red fluorescence. (C) Live imaging of SH-SY5Y cells allowed observation of GDNF distribution among the cells.
- Live cell imaging of recombinant human GDNF-Biotin protein in live intact human SH-SY5Y cells.(A) CellMask™ Actin 1X solution was applied for 30 minutes at 370C, resulting in a green fluorescence to visualize cellular membrane. (B) Following this, the same cells underwent incubation with 0.1 µM of recombinant human GDNF protein-Biotin (#G-240-B) for 30 minutes at 370C, followed by Alexa Streptavidin-647, leading to red fluorescence. (C) Live imaging of SH-SY5Y cells allowed observation of GDNF distribution among the cells.
- human GDNF-Biotin transport along the MN axons.human GDNF-Biotin (#G-240-B) is transported retrogradely in a MN explant grown in a microfluidic chamber. Representative images from a 10’ time lapse, taken roughly 2 hours after 50 ng/ml QDot-GDNF addition to distal axon compartment. Arrowheads point at transported particles. Live imaging was performed using a 60x objective with spinning disc confocal microscope, with a 2000ms interval between frames. Horizontal scale =10μm, vertical scale = 50 seconds.
- Indirect flow cytometry of recombinant human GDNF protein in live intact human SH-SY5Y cells.___ SH-SY5Y cells.___ SH-SY5Y cells + 100 nM recombinant human GDNF protein-Biotin (#G-240-B) + 2 µg/ml Streptavidin, Alexa Fluor™ 647 conjugate.
___ SH-SY5Y cells + 100 nM recombinant human GDNF protein (#G-240) + 2 µg/ml Streptavidin, Alexa Fluor™ 647 conjugate.
___ SH-SY5Y cells + 2 µg/ml Streptavidin, Alexa Fluor™ 647 conjugate. - Recombinant human GDNF-Biotin protein induces ERK1/2 MAPK activation in SH-SY5Y cells.Cells were serum starved for 2 h and then stimulated with various concentrations of recombinant human GDNF-Biotin protein (#G-240-B) for 15 min. Cell proteins were resolved by SDS-PAGE and probed with anti-phospho-ERK1/2 (lower panel) and anti-ERK1/2 (upper panel).
Glial-derived neurotrophic factor (GDNF) is a member of the TGF-β superfamily. GDNF signals through a multi-component receptor system, composed of a RET proto-oncogene and one of the four α1-α4 receptors.1
GDNF promotes survival of various neuronal cells, including motoneurons,2,3 Purkinje cells and sympathetic neurons.4 In embryonic midbrain cultures, GDNF promotes the survival and morphological differentiation of dopaminergic neurons and increases their high-affinity DA uptake.5 Cells that express GDNF include Sertoli cells, type 1 astrocytes, Schwann cells,6 neurons, pinealocytes, and skeletal muscle cells.7
In vivo, following transection of facial motor neuron axons, locally applied GDNF has been shown to rescue virtually all damaged neurons from cell death.8 GDNF may be of clinical relevance in the treatment of Parkinson's disease that is characterized by progressive degeneration of midbrain dopaminergic neurons.9,10