Name: Recombinant mouse proBDNF (cleavage resistant) protein
Brand: Alomone Labs
SKU: B-243

Cat #: B-243

Alternative Name proBrain-Derived Neurotrophic Factor

Lyophilized Powder yes

Bioassay Tested

Sterile & Endotoxin Free yes

Origin Recombinant, E. coli

MW: 52 kDa

Endotoxin Level <0.1 EU per 1 µg of the protein by the LAL method.

Purity: >98% (HPLC)

Form Lyophilized from a 0.2 µm filtered solution.

Effective concentration 0.2 nM.

Sequence MAPMKEVNVHGQGNLAYPGVRTHGTLESVNGPRAGSRGLTTTSLADTFEHVIEELLDEDQKVRPNEENHKDADLYTSRVMLSSQVPLEPPLLFLLEEYKNYLDAANMSMRVAGHSDPARRGELSVCDSISEWVTAADKKTAVDMSGGTVTVLEKVPVSKGQLKQYFYETKCNPMGYTKEGCRGIDKRHWNSQCRTTQSYVRALTMDSKKRIGWRFIRIDTSCVCTLTIKRGR (shown in bold is the prodomain, underlined are the double point mutations preventing cleavage).

Structure

Activity BDNF is a neurotrophic factor and binds p75^NTR as well as TrkB receptors^1,2. BDNF supports the survival of many cell types^3-8. proBDNF has been shown to be a pro-apoptotic ligand for sympathetic neurons⁹ expressing both p75 and sortilin, and to be involved in LTD¹⁰.

References-Activity

Tolkovsky, A. (1997) Trends Neurosci. 20, 1.
Jing, S. et al. (1992) Neuron 9, 1067.
Acheson, A. et al. (1995) Nature 374, 450.
Morse, J.K. et al. (1993) J. Neurosci. 13, 4146.
Hyman, C. et al. (1991) Nature 350, 230.
Friedman, B. et al. (1995) J. Neurosci. 15, 1044.
Meyer, M. et al. (1992) J. Cell Biol. 119, 45.
Koliatsos, V.E. et al. (1993) Neuron 10, 359.
Teng, H.K. et al. (2005) J. Neurosci. 25, 5455.
Woo, N.H. et al. (2005) Nat. Neurosci. 8, 1069.

Shipping and storage Shipped at room temperature. Product as supplied can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.

Solubility Sterile water at a concentration of at least 10 µg/ml.
Centrifuge all product preparations before use (10000 x g 5 min). Repeated freezing/thawing might result in loss of activity.

Storage of solutions Up to one week at 4°C or four-six weeks at -70°C.

Our bioassay

Alomone Labs Recombinant mouse proBDNF (cleavage resistant) protein mediates neurite outgrowth in TrkB transfected PC12 cells.
One day post transfection, the cells were stimulated with 0.2 nM Recombinant human BDNF protein (#B-250), 0.2 nM Recombinant mouse proBDNF protein (#B-240) and 0.2 nM Recombinant mouse proBDNF (cleavage resistant) protein (#B-243). Development of neurites was visualized by fluorescence microscopy after 4 days.
Alomone Labs Recombinant mouse proBDNF (cleavage resistant) protein mediates ERK1/2 activation in TrkB transfected HEK-293 cells.
Three days post transfection, cells were serum-starved for 2h and then challenged for 10 min with Recombinant human BDNF protein (#B-250), Recombinant mouse proBDNF protein (#B-240) or Recombinant mouse proBDNF(cleavage resistant) protein (#B-243). Cell proteins were resolved by SDS PAGE and the level of ERK1/2 phosphorylation determined using anti-phospho ERK.

Scientific background

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.⁵

The 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.⁹

In 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.

Peptide Content: 100%

Lyophilized Powder

Last update: 06/11/2022

For research purposes only, not for human use

Specifications

Citations

Applications

Scientific Background

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Certificate of Analysis

Recombinant mouse proBDNF (cleavage resistant) protein

Specifications

Citations

Applications

Scientific Background

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