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Home / Proteins & Peptide / Proteins

Recombinant human Val66Met proBDNF (cleavage resistant) protein

SKU : BHP21300021

A Neurotrophic Factor

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Mfr.No.	B-456, B-456_1 mcg, B-456_10 mcg, B-456_5 mcg
Description	A Neurotrophic Factor
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	Brain-derived neurotrophic factor (BDNF) is a neurotrophic factor that bind p75NTR as well as TrkB receptors1,2. BDNF supports the survival of many cell types3-8 and also modulates hippocampal plasticity and hippocampal-dependent memory in cell models and in animals9.The BDNF gene, like other peptide growth factors, encodes a precursor peptide (proBDNF), which is proteolytically cleaved to form the mature protein10. proBDNF has been shown to be a pro-apoptotic ligand for sympathetic neurons expressing both p75NTR and sortilin11, and to be involved in long-term potentiation (LTP) stage of the memory-related modifications in synaptic transmission12.A nonconservative single nucleotide polymorphism (SNP) in the human BDNF gene has been identified at nucleotide 196 (G/A) producing an amino acid substitution (Valine to Methionine) at codon 66 (Val66Met, rs 6265). Although located in the 5' pro-BDNF region, this SNP resulted in striking deficits in the cellular distribution and regulated secretion of the mature protein, BDNF and hence in corresponding alterations of human hippocampal function and episodic memory in vivo9.Egan MF. et al demonstrated the molecular mechanisms that control activity-dependent BDNF secretion and showed that depolarization-dependent secretion of BDNF in hippocampal neurons is significantly impaired when this Val66Met SNP occurs. Using double-staining techniques, they demonstrated that Val-BDNF-containing secretory granules are colocalized with synaptophysin, a marker for synapses. In contrast, Val66Met-BDNF aggregates are accumulated in the cell body and rarely colocalize with synaptophysin. This suggests that even if it can be secreted in small amounts near the cell body through the constitutive pathway, the Met-BDNF protein cannot be secreted at synapses9. Studies of heterozygote BDNF knockout rodents, who presumably have intermediate BDNF levels, demonstrate clear physiological13 and behavioral14 abnormalities, suggesting that secretion levels are critical.Multiple studies over the recent decades in humans, in vivo in animal models and in vitro found an association between the Val66Met polymorphism and bipolar and unipolar disorders15, Schizophrenia16,17, anxiety-related behavior18,19 and controversial association with ADHD20,21.The data emerged from the analysis of the Val66Met phenotype in various syndromes and diseases reveal the importance of the pro-region of the BDNF polypeptide, particular Valine66 and perhaps the nearby sequence, in intracellular trafficking and secretion of BDNF.
Biological Activity	Brain-derived neurotrophic factor (BDNF) is a neurotrophic factor that bind p75NTR as well as TrkB receptors1,2 and supports the survival of many cell types3-8. proBDNF has been shown to be a pro-apoptotic ligand for sympathetic neurons9 expressing both p75NTR and sortilin, and to be involved in long-term potentiation (LTP) stage of the memory-related modifications in synaptic transmission10.
Formulation	Lyophilized from double distilled water (ddH2O). May contain TFA as a residual counter ion.
Molecular Weight	51.45 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	Val66Met proBrain-Derived Neurotrophic Factor (Mut-human)
Gene Id	NGFR,NTRK2,SORT1,SORCS2
Sequence	MAPMKEANIRGQGGLAYPGVRTHGTLESVNGPKAGSRGLTSLADTFEHMIEELLDEDQKVRPNEENNKDADLYTSRVMLSSQVPLEPPLLFLLEEYKNYLDAANMSMRVAGHSDPARRGELSVCDSISEWVTAADKKTAVDMSGGTVTVLEKVPVSKGQLKQYFYETKCNPMGYTKEGCRGIDKRHWNSQCRTTQSYVRALTMDSKKRIGWRFIRIDTSCVCTLTIKRGR (the prodomain is shown in bold, underlined are the double point mutations preventing cleavage (AG). The Val66Met mutation is underlined in the sequence).
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

Reference	Tolkovsky, A. (1997) Trends Neurosi. 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. Egan, M.F. et al. (2003) Cell 112, 257. Seidah, N.G. et al. (1996) FEBS Lett. 379, 247. Teng, H.K. et al. (2005) J. Neurosci. 25, 5455. Woo, N.H. et al. (2005) Nat. Neurosci. 8, 1069. Korte, M. et al. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 8856. Lyons, W.E. et al. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 15239. Post, R.M. (2007) J. Psychiatric Res. 41, 979. Ho, B.C. et al. (2006) Arch. Gen. Psychiatry 63, 731. Rosa, A. et al. (2006) Am. J. Med. Gen. B Neuropsych. Gen. 141B, 135. Hashimoto, K. (2007) BioEssays 29, 116. Chen Z.Y. et al. (2006) Science 314, 140. Kent, L. et al. (2005) Mol. Psychiatry 10, 939. Sanchez-Mora C. et al. (2010) Am. J. Med. Gen. B Neuropsych. Gen. 153B, 512. Tolkovsky, A. (1997) Trends Neurosi. 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.

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Combination SKU	Supplier No.	Size	Your price	Quantity
BHP21300021-1	B-456_1 mcg	1 mcg	$261		Add to Cart
BHP21300021-2	B-456_10 mcg	10 mcg	$1268		Add to Cart
BHP21300021-3	B-456_5 mcg	5 mcg	$705		Add to Cart
BHP21300021-4	B-456_1 mcg	5x1 mcg	$1030		Add to Cart
BHP21300021-5	B-456_5 mcg	5x5 mcg	$2811		Add to Cart

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