recombinant human probdnf  (Alomone Labs)

Journal: The Journal of Neuroscience

Article Title: SPIG1 Negatively Regulates BDNF Maturation

doi: 10.1523/JNEUROSCI.1597-13.2014

Figure Lengend Snippet: Effect of SPIG1 on BDNF expression in PC12 cells. A , Colocalization of SPIG1 and proBDNF in vesicle-like structures. Twelve hours after transfection with the indicated constructs, PC12 cells were differentiated with NGF as described in Materials and Methods.

Article Snippet: A 96-well polystyrene ELISA plate (#9018, Costar) was coated with 4 pmol of purified recombinant human proBDNF (B-257, Alomone Labs) or mature BDNF (GF029, Millipore).

Techniques: Expressing, Transfection, Construct

Journal: The Journal of Neuroscience

Article Title: SPIG1 Negatively Regulates BDNF Maturation

doi: 10.1523/JNEUROSCI.1597-13.2014

Figure Lengend Snippet: A mechanism model for the position-specific branching of RGC axons. Both ephrin-A/EphA signaling (light yellow) in the distal part and proBDNF/ephrin-A-p75 NTR signaling (yellow) in the proximal part have been shown to mediate the suppression of branching

Article Snippet: A 96-well polystyrene ELISA plate (#9018, Costar) was coated with 4 pmol of purified recombinant human proBDNF (B-257, Alomone Labs) or mature BDNF (GF029, Millipore).

Techniques:

Journal: The Journal of Neuroscience

Article Title: SPIG1 Negatively Regulates BDNF Maturation

doi: 10.1523/JNEUROSCI.1597-13.2014

Figure Lengend Snippet: Subcellular colocalization of exogenous SPIG1 and proBDNF in the chick RGC. A , Colocalization of SPIG1 and proBDNF in axon terminals. After the electroporation of SPIG1 (FLAG-tagged at the C terminus) and BDNF constructs at HH stage 9, retinal cells were

Article Snippet: A 96-well polystyrene ELISA plate (#9018, Costar) was coated with 4 pmol of purified recombinant human proBDNF (B-257, Alomone Labs) or mature BDNF (GF029, Millipore).

Techniques: Electroporation, Construct

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: Exogenous proBDNF induces pain hypersensitivity and spinal cord activation in mice. ( A ) Dosage effect of exogenous proBDNF protein on PWT by injection of proBDNF protein into the plantar (*P

Article Snippet: Other recombinant human proBDNF, mice proBDNF and human mBDNF for ELISA, Western Blot or behavior studies were purchased from Alomone Labs (Israel).

Techniques: Activation Assay, Mouse Assay, Injection

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: Polyclonal Ab-proBDNF pretreatment attenuates inflammatory pain in mice. ( A,B ) proBDNF polyclonal antibody (5 ml/Kg) i.p pretreatment attenuated both phases of nociceptive responses induced by 5% formalin intra-plantar injection in Kunming mice. ( A ) Time course of biphasic nociceptive response (*P

Article Snippet: Other recombinant human proBDNF, mice proBDNF and human mBDNF for ELISA, Western Blot or behavior studies were purchased from Alomone Labs (Israel).

Techniques: Mouse Assay, Injection

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: Characterization of proBDNF monoclonal antibody 2B11. ( A ) ELISA assay for the immunoreactivity of 2B11 against human proBDNF prodomain, and human, rat and mice proBDNF proteins, and human mature BDNF (mBDNF). 2B11 has strong immunoreactivity against proBDNF and prodomain, but not mBDNF; ( B ) Representative Western blot of human proBDNF and mBDNF detected by 2B11 (dilution 1:2000), note that 2B11 specifically recognizes proBDNF, but not mBDNF. ( C ) Representative images of neurosphere radiant migration treated by proBDNF, mBDNF, sheep polyclonal anti-proBDNF antibody, mouse monoclonal anti-proBDNF antibody 2B11 and co-treatment. ( D ) Statistical analysis of neurosphere migration radiance assay (***P

Article Snippet: Other recombinant human proBDNF, mice proBDNF and human mBDNF for ELISA, Western Blot or behavior studies were purchased from Alomone Labs (Israel).

Techniques: Enzyme-linked Immunosorbent Assay, Mouse Assay, Western Blot, Migration

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: Upregulation of p75NTR and effect of local proBDNF injection on inflammatory reaction in mice. ( A ) Representative Western blot of p75NTR and sortilin ( a ) and the semi-quantitative analysis of their expression (b and c) after formalin intra-plantar injection (***p

Article Snippet: Other recombinant human proBDNF, mice proBDNF and human mBDNF for ELISA, Western Blot or behavior studies were purchased from Alomone Labs (Israel).

Techniques: Injection, Mouse Assay, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Peripheral Brain Derived Neurotrophic Factor Precursor Regulates Pain as an Inflammatory Mediator

doi: 10.1038/srep27171

Figure Lengend Snippet: Upregulation of proBDNF in the local tissue in acute and persistent inflammatory pain in mice. ( A ) Representative Western blot (a) and their semi-quantitative analyses of mature BDNF (b), proBDNF (c) and their ratio (d) in the local tissue after 10 μL 5% formalin intra-plantar injection into Kunming mice (*p

Article Snippet: Other recombinant human proBDNF, mice proBDNF and human mBDNF for ELISA, Western Blot or behavior studies were purchased from Alomone Labs (Israel).

Techniques: Mouse Assay, Western Blot, Injection

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Activation Regulates the Timing of the Maturation of Cortical Parvalbumin Interneuron Connectivity and Promotes Juvenile-like Plasticity in Adult Visual Cortex

doi: 10.1523/JNEUROSCI.2881-18.2019

Figure Lengend Snippet: proBNDF-mediated p75NTR activation in cortical PV cells reduces their perisomatic boutons. A , Experimental approach. B , The intensity of perisomatic PV immunostaining (green) is reduced in the binocular visual cortex ipsilateral to the minipump-releasing mut-proBDNF (Ipsi) compared with the contralateral cortex (Contra) in the same animal. On the other hand, perisomatic PV intensity in the ipsilateral cortex of PV_Cre;p75 flx/flx mice is similar to that observed in the contralateral, untreated cortex. C , Low ( C1 ) and high ( C2 ) magnification of PNN (red, WFA staining) enwrapping PV cells (green) shows a dramatic reduction in both PNN density and intensity in the visual cortex infused with mut-proBFNF. This effect is abolished in PV_Cre;p75 flx/flx mice. Scale bars: C1 , 100 μm; B , C2 , 10 μm. D , Quantification of the mean intensity of perisomatic PV-positive puncta in ipsilateral compared with contralateral cortex. I/C ratio is obtained for each animal and then averaged between different animals. Mean I/C ratio is significantly reduced in Mut-proBDNF-infused p75 Ctrl mice compared with Mut-proBDNF-infused PV_Cre;p75 flx/flx mice (unpaired t test, df = 8, t = 6.077, p = 0.0003). E , The ratio of mean PNN intensity around PV cells in ipsilateral versus contralateral cortex is significantly lower in p75 Ctrl than PV_Cre;p75 flx/flx mice infused with mut-proBDNF (unpaired t test, df = 8, t = 15.33, p

Article Snippet: Minipumps (model 1007D; flow rate 0.5 μl/h; Alzet) were filled with mut-proBDNF (1 μg/ml in filtered PBS, Alomone Labs) or vehicle solution and connected to a cannula (gauge 30) implanted directly in the primary visual cortex (2.5 mm lateral to the midline, 2.5 mm anterior to λ).

Techniques: Activation Assay, Immunostaining, Mouse Assay, Staining

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Activation Regulates the Timing of the Maturation of Cortical Parvalbumin Interneuron Connectivity and Promotes Juvenile-like Plasticity in Adult Visual Cortex

doi: 10.1523/JNEUROSCI.2881-18.2019

Figure Lengend Snippet: Modulation of tPA activity affects the formation of PV cell innervations during early postnatal development. A , Control EP18 PV cell ( A1 , green represents Ctrl). B , PV cell treated with the tPA inhibitor PPACK from EP10–EP18 shows simpler axonal arborization, contacting less potential targets ( B2 , blue represents NeuN-positive somata). C , PV cell treated with tPA in the same time window shows a very complex axonal arbor ( C2 ) and an increase in both terminal branching and perisomatic boutons ( C3 , arrowheads) compared with control cells ( A2 , A3 ). D , PV cell treated simultaneously with tPA and mut-proBDNF shows axonal branching and perisomatic innervation more similar to those formed by PV cell treated with mut-proBDNF alone, suggesting that the effects of tPA application may be mediated by a decrease in endogenous proBDNF/mBDNF ratio. Stars indicate NeuN-positive somata that are not innervated. Scale bars: A1–D1 , 50 μm; A2–D2 , 10 μm; A3–D3 , 5 μm. E , Perisomatic boutons density (one-way ANOVA, F (3,20) = 121.2, p

Article Snippet: Minipumps (model 1007D; flow rate 0.5 μl/h; Alzet) were filled with mut-proBDNF (1 μg/ml in filtered PBS, Alomone Labs) or vehicle solution and connected to a cannula (gauge 30) implanted directly in the primary visual cortex (2.5 mm lateral to the midline, 2.5 mm anterior to λ).

Techniques: Activity Assay

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Activation Regulates the Timing of the Maturation of Cortical Parvalbumin Interneuron Connectivity and Promotes Juvenile-like Plasticity in Adult Visual Cortex

doi: 10.1523/JNEUROSCI.2881-18.2019

Figure Lengend Snippet: proBNDF-mediated p75NTR activation in cortical PV cells restores ocular dominance plasticity in adult visual cortex in vivo . A , Typical VEP responses to the stimulation of either contralateral (blue) or ipsilateral (red) eye to the cortex in which the recording is performed in p75NTR Ctrl mice infused with either vehicle or mut-proBDNF, and PV_Cre;p75NTR flx/flx mice infused with mut-proBDNF. Calibration bars: 50 μV, 100 ms. B , C/I VEP ratio mean values. Three days of monocular deprivation do not affect the C/I VEP ratio in adult mice, although it leads to a significant decrease in the C/I VEP ratio in animals treated with mut-proBDNF. Mut-proBDNF effects are, however, abolished in PV_Cre;p75 flx/flx mice (one-way ANOVA, F (2,18) = 8.903, p = 0.0021). p75NTR Ctrl + vehicle: n = 9 mice; p75NTR Ctrl + mut-proBDNF: n = 5 mice; PV_Cre;p75 flx/flx +mut-proBDNF: n = 7 mice. C , ODI of p75NTR Ctrl mice infused with vehicle solution and PV_Cre;p75 flx/flx mice infused with mut-proBDNF are not significantly different from those of undeprived animals, whereas ODIs in p75 Ctrl mice treated with mut-proBDNF are significantly shifted toward the open eye (one-way ANOVA, F (2,443) = 5.203, p = 0.0058). D , Mean spontaneous discharge is significantly increased only in p75 Ctrl mice treated with mut-proBDNF (one-way ANOVA, F (2,443) = 4.580, p = 0.0107). p75NTR Ctrl + vehicle: n = 9 mice, 174 cells; p75NTR Ctrl + mut-proBDNF: n = 5 mice, 147 cells; PV_Cre;p75 flx/flx +mut-proBDNF: n = 6 mice, 125 cells. Gray area represents the C/I VEP ratio ( B ) or the ODI range ( C ) (mean ± SEM) in adult nondeprived animals ( n = 5 mice, 99 cells). * indicate p

Article Snippet: Minipumps (model 1007D; flow rate 0.5 μl/h; Alzet) were filled with mut-proBDNF (1 μg/ml in filtered PBS, Alomone Labs) or vehicle solution and connected to a cannula (gauge 30) implanted directly in the primary visual cortex (2.5 mm lateral to the midline, 2.5 mm anterior to λ).

Techniques: Activation Assay, In Vivo, Mouse Assay, Mass Spectrometry

Journal: The Journal of Neuroscience

Article Title: p75 Neurotrophin Receptor Activation Regulates the Timing of the Maturation of Cortical Parvalbumin Interneuron Connectivity and Promotes Juvenile-like Plasticity in Adult Visual Cortex

doi: 10.1523/JNEUROSCI.2881-18.2019

Figure Lengend Snippet: mut-proBDNF destabilizes PV cell innervation, even after it has reached maturity. A , Control PV cell ( A1 , Ctrl, green) at EP32 with exuberant innervation field characterized by extensive branching contacting the majority of potential targets, dense boutons along axons ( A2 ), and terminal branches with prominent and clustered boutons ( A3 ; arrowheads) around NeuN-positive somata (blue). B , PV cell treated with wt-proBDNF from EP26-EP32 shows overall similar axon size ( B1 ), percentage of potentially targeted neurons ( B2 ), and perisomatic innervations ( B3 ) as control, untreated PV cells. C , PV cell treated with mut-proBDNF from EP26-EP32 shows a drastic reduction both in percentage of innervated cells ( C2 ) and perisomatic innervation ( C3 ). Stars indicate NeuN-positive somata that are not innervated. Scale bars: A1–C1 , 50 μm; A2–C2 , 10 μm; A3–C3 , 5 μm. D , Perisomatic bouton density (one-way ANOVA, F (2,18) = 93.34, p

Article Snippet: Minipumps (model 1007D; flow rate 0.5 μl/h; Alzet) were filled with mut-proBDNF (1 μg/ml in filtered PBS, Alomone Labs) or vehicle solution and connected to a cannula (gauge 30) implanted directly in the primary visual cortex (2.5 mm lateral to the midline, 2.5 mm anterior to λ).

Techniques:

Journal: Translational Psychiatry

Article Title: BDNF–TrkB signaling in the nucleus accumbens shell of mice has key role in methamphetamine withdrawal symptoms

doi: 10.1038/tp.2015.157

Figure Lengend Snippet: Depression-like behavior and levels of proBDNF and BDNF in the brain regions after withdrawal from repeated METH exposure. ( a ) Schedule of treatment and behavioral tests. Saline (10 ml kg −1 per day for 5 days) or METH (3 mg kg −1 per day for 5 days) was injected into mice. Behavioral tests were performed at days 7, 11 and 18 (SPT), and days 8, 12 and 19 (LMT, TST, FST). ( b ) SPT: sucrose preference of METH-treated mice was significantly lower than that of control (saline-treated) mice. LMT: there were no differences between control and METH-treated mice. TST and FST: the immobility time of METH-treated mice was significantly higher than that of control mice. * P

Article Snippet: The following primary antibody was used: proBDNF (1:400, Alomone Labs, Jerusalem, Israel), BDNF (1:200, H-117, Santa Cruz Biotechnology, Danvers, CA, USA), phosphorylated-TrkB (Tyr 706) (1:200, Santa Cruz Biotechnology), TrkB (80E3) (1:1000, Cell Signaling Technology, Danvers, MA, USA), GluA1 (1 μg ml−1 , Abcam, Cambridge, UK) and β-actin (1:10 000, Sigma-Aldrich).

Techniques: Injection, Mouse Assay, Single-particle Tracking