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Recombinant human proBDNF (cleavage resistant) protein

SKU : BHP21300014

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

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Mfr.No.	B-256, B-256_0.1 mg, B-256_0.25 mg, B-256_0.5 mg, B-256_1 mg, B-256_10 mcg, B-256_2 mcg, B-256_25 mcg, B-256_5 mcg, B-256_50 mcg
Description	Human proBrain-Derived Neurotrophic Factor (cleavage resistant), Recombinant, E. coli
Reconstitution	Centrifuge the vial (10,000 x 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.
Background	BDNF is a neurotrophic factor produced by proteolytic cleavage of its precursor, proBDNF.1 The actions of BDNF are mediated via the binding to TrkB or p75.2,3The precursor form 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 p75, has ushered in a new era which reexamines the biological roles of the two forms.6 Some 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 ERK1/2.9In brain homogenates a mixture of both, proBDNF and mature BDNF has been found10,11 and in cortical neurons secretion of proBDNF has been shown.7 Binding of both proBDNF and mature BDNF to TrkB has been proposed to be via the R103 residue in the mature portion.9
Biological Activity	BDNF is a neurotrophic factor and binds p75NTR as well as TrkB receptors1,2. BDNF supports the survival of many cell types3-8. proBDNF has been shown to be a pro-apoptotic ligand for sympathetic neurons9 expressing both p75 and sortilin, and to be involved in LTD10.
Formulation	Lyophilized from filtrated Ammonium acetate solution. May contain acetate as a residual counter ion.
Molecular Weight	51.41 kDa (dimer)
Solubility	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.
Source	Recombinant, E. coli
Concentration	0.1 - 10 nM
Form	Lyophilized
Notes	Bioassay Tested: yes Steril endotoxin free: yes Is Toxin: No
UNSPSC Code	12352202
Lead Time	1-2 Business Days

Synonyms	proBrain-Derived Neurotrophic Factor
Gene Id	NGFR,NTRK2,SORT1,SORCS2
Sequence	MAPMKEANIRGQGGLAYPGVRTHGTLESVNGPKAGSRGLTSLADTFEHVIEELLDEDQKVRPNEENNKDADLYTSRVMLSSQVPLEPPLLFLLEEYKNYLDAANMSMRVAGHSDPARRGELSVCDSISEWVTAADKKTAVDMSGGTVTVLEKVPVSKGQLKQYFYETKCNPMGYTKEGCRGIDKRHWNSQCRTTQSYVRALTMDSKKRIGWRFIRIDTSCVCTLTIKRGR (the prodomain is shown in bold, underlined are the double point mutations preventing cleavage)
Target	p75NTR, Sortilin receptors

Country Of Origin	Israel/IL
Purity	>=98% (HPLC)
Storage	Storage After Reconstitution: The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods (up to 6 months), small aliquots should be stored at -20°C. We do not recommend storing the product in working solutions for longer than a few days. Avoid multiple freeze-thaw cycles. Storage Before Reconstitution: The product is shipped as a lyophilized powder at room temperature. Upon receipt, store the product at -20°C. Protect from moisture. Storage of solutions: The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods (up to 6 months), small aliquots should be stored at -20°C. We do not recommend storing the product in working solutions for longer than a few days. Avoid multiple freeze-thaw cycles.

Applications	Western blot, Neurite outgrowth assay

Reference	Cahoon-Metzger, S.M. et al. (2001) Dev. Biol. 232, 246. Troy, C.M. et al. (2002) J. Biol. Chem. 277, 34295. Chen, Z.Y. et al. (2004) J. Neurosci. 24, 4401. Egan, M.F. et al. (2003) Cell 112, 257. Lee, R. et al. (2001) Science 294, 1945. Teng, H.K. et al. (2005) J. Neurosci. 25, 5455. Woo, N.H. et al. (2005) Nat. Neurosci. 8, 1069. Fayard, B. et al. (2005) J. Neurosci. Res. 80, 18. Michalski, B. et al. (2003) Mol. Brain Res. 111, 148. Zhou, X.F. et al. (2004) J. Neurochem. 91, 704. Bibel, M. and Barde, Y.A. (2000) Genes Dev. 14, 2919. 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. Tolkovsky, A. (1997) Trends Neurosci. 20, 1.

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Combination SKU	Supplier No.	Size	Your price	Quantity
BHP21300014-1	B-256_0.1 mg	0.1 mg	$1595		Add to Cart
BHP21300014-2	B-256_0.25 mg	0.25 mg	$2839		Add to Cart
BHP21300014-3	B-256_0.5 mg	0.5 mg	$4515		Add to Cart
BHP21300014-4	B-256_1 mg	1 mg	$5524		Add to Cart
BHP21300014-5	B-256_10 mcg	10 mcg	$210		Add to Cart
BHP21300014-6	B-256_2 mcg	2 mcg	$76		Add to Cart
BHP21300014-7	B-256_25 mcg	25 mcg	$479		Add to Cart
BHP21300014-8	B-256_5 mcg	5 mcg	$170		Add to Cart
BHP21300014-9	B-256_50 mcg	50 mcg	$845		Add to Cart

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