mouse proBDNF (cleavage resistant)

Western blot, Neurite outgrowth assay

p75NTR, Sortilin receptors

The product is shipped as a lyophilized powder at room temperature. Upon receipt, store the product at -20°C. Protect from moisture.

52 kDa (dimer)

Mouse proBrain-Derived Neurotrophic Factor (cleavage resistant), Recombinant, E. coli

Centrifuge the vial (10, 000 × g for 5 minutes) before adding solvent to spin down all the powder to the bottom of the vial. The lyophilized product may be difficult to visualize. Add solvent directly to the centrifuged vial. Gently tap, tilt, and roll the vial to aid dissolution. Avoid vigorous vortexing; light vortexing for up to 3 seconds is acceptable if needed. 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. Repeated freeze-thaw cycles may result in loss of activity.

7, 4

0.2 nM

Centrifuge the vial before adding solvent (10, 000 x g for 5 minutes) to spin down all the powder to the bottom of the vial. The lyophilized product may be difficult to visualize. Add solvent directly to the centrifuged vial. Tap the vial to aid in dissolving the lyophilized product. Tilt and gently roll the liquid over the walls of the vial. Avoid vigorous vortexing. Light vortexing for up to 3 seconds is acceptable if needed. 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.

yes

BDNF is a neurotrophic factor produced by proteolytic cleavage of its precursor, proBDNF. The biologically relevant form of the protein was thought to be the mature form, BDNF, which has been shown to affect the development of motoneurons, modulate synaptic transmission and affect axonal branching, dendrite growth and synapse number.1 These actions are mediated via the binding of BDNF to TrkB. Different actions are mediated by the binding of BDNF to p75, such as neuronal process retraction2 and neuronal apoptosis.3 The precursor form of the neurotrophic factor was thought to be important for the correct folding, secretion and trafficking of the mature protein. A single-nucleotide polymorphism (Val66 to Met) in the pro-domain of the human BDNF gene impairs intracellular trafficking and regulated secretion of BDNF in primary cortical neurons and neurosecretory cells but not in endothelial and vascular cells.4 This has been shown to affect memory and lead to abnormal hippocampal function in humans.5The finding that proBDNF and not mature BDNF is the preferred ligand for p756 has ushered in a new era which reexamines the biological roles of the two forms. Contradictory biological roles for proBDNF have been proposed. It has been shown to be a pro-apoptotic ligand for sympathetic neurons7 expressing both p75 and sortlin, and to be involved in LTD8. On the other hand, it has also been shown to elicit prototypical TrkB responses in biological assays, such as TrkB tyrosine phosphorylation, and activation of ERK1/2.9 Binding of both proBDNF and mature BDNF to TrkB has been proposed to be via the R103 residue in the mature portion.9In addition, the question of the relative abundance of the precursor versus the mature form has been investigated. In brain homogenates a mixture of proBDNF and mature BDNF has been found.10, 11 The secretion of proBDNF from cortical neurons has been shown.7 Most of the work showing secretion of proBDNF was done in cell lines overexpressing full length BDNF constructs.4, 6, 12, 13 It has been argued14 that this model misrepresents the authentic situation in vivo, since in the transfected systems the secretion of proBDNF results from overloading the limited capacity of the processing machinery. These authors argue that in fact proBDNF is a transient intermediate and is rapidly converted intracellularly to mature BDNF.