recombinant mouse cleavage resistant probdnf  (Alomone Labs)

Journal: bioRxiv

Article Title: CNTF specifically slows down the axonal transport of signalling endosomes

doi: 10.1101/2025.10.09.681259

Figure Lengend Snippet: A) Male and female ChAT.eGFP mice were allocated into one of five groups: i) PBS (control); ii) CNTF; iii) proBDNF; iv) HGF; and v) NTRN. B) Each mouse received intramuscular injections of H C T combined with a single NTF into the left tibialis anterior and right soleus muscles to target fast and slow motor neurons, respectively. After a 4-8 h incubation period, time-lapse microscopy was performed on both sciatic nerves. C) H C T-labelled signalling endosomes (pseudo-coloured in magenta) from single ChAT.eGFP motor axons were individually tracked to quantify retrograde transport dynamics. Three representative retrogradely transported signalling endosomes are identified by yellow, cyan, and peach arrowheads connected by dashed lines across frames. Grey arrowheads and dashed lines identify a stationary endosome. See also Video 1 . Scale bar = 5 μm, frame interval = 3 s.

Article Snippet: Mice were divided into five experimental cohorts, and received 5-7 μg of H C T mixed with: a) phosphate buffered saline (PBS) as the control; b) 50 ng recombinant human CNTF protein (Peprotech, 450-13), c) 50 ng recombinant mouse cleavage-resistant proBDNF (Alomone labs, B-243), d) 25 ng recombinant human HGF (Bio-Techne, 294-HG[CF]), or e) 50 ng recombinant human NTRN (Peprotech, 450-11), as summarised in .

Techniques: Control, Muscles, Incubation, Time-lapse Microscopy

Journal: bioRxiv

Article Title: CNTF specifically slows down the axonal transport of signalling endosomes

doi: 10.1101/2025.10.09.681259

Figure Lengend Snippet: In both fast motor neurons (FMNs) and slow motor neurons (SMNs), proBDNF did not alter A) mean endosome speed ( p = 0.06 for motor neuron type; p = 0.19 for stimulation factor; p = 0.70 for interaction), B) maximum endosome speed ( p = 0.30 for motor neuron type; p = 0.62 for stimulation factor; p = 0.25 for interaction) or C) pausing percentage ( p = 0.24 for motor neuron type; p = 0.92 for stimulation factor; p = 0.34 for interaction). D ) Violin plots of individual endosomes show comparable distributions in all conditions (FMNs: PBS mean = 2.78 µm/s ± 0.03 [n = 495], proBDNF mean = 2.67 µm/s ± 0.03 [n = 496]; SMNs: PBS mean = 2.57 µm/ s ± 0.03 [n = 495], proBDNF mean = 2.53 µm/s ± 0.03 [n = 477]). Overlapping endosome frame-to-frame and mean endosome speed distribution curves confirm that proBDNF does not modulate retrograde transport in E) FMNs or F) SMNs. Statistical analyses were performed using two-way ANOVA and Holm-Šídák 1 s multiple comparisons tests. ns, not significant. n = 7-8. Black circles = males (n = 4 PBS; n = 2 CNTF), white circles = females (n = 4 PBS; n = 5 proBDNF).

Article Snippet: Mice were divided into five experimental cohorts, and received 5-7 μg of H C T mixed with: a) phosphate buffered saline (PBS) as the control; b) 50 ng recombinant human CNTF protein (Peprotech, 450-13), c) 50 ng recombinant mouse cleavage-resistant proBDNF (Alomone labs, B-243), d) 25 ng recombinant human HGF (Bio-Techne, 294-HG[CF]), or e) 50 ng recombinant human NTRN (Peprotech, 450-11), as summarised in .

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Maternal immune activation-induced proBDNF-mediated neural information processing dysfunction at hippocampal CA3-CA1 synapses associated with memory deficits in offspring

doi: 10.3389/fcell.2022.1018586

Figure Lengend Snippet: The performance in the conditioning and fear memory test. PolyI:C-treated offspring were intra -CA1 or -CA3 infused with anti-proBDNF antibody or p75 NTR inhibitor TAT-Pep5 30 min before the testing. The freezing levels of polyI:C-treated offspring during the training (A) and the memory test (B) are lower while blocking the activation of proBDNF/p75 NTR signaling can effectively reversed the impaired memory behavior. Data are presented as mean ± SEM. * p < 0.05, Control + ACSFCA1 vs. other groups in A and the group vs. Control + ACSFCA1, PolyI:C + AntiCA1 and PolyI:C + Pep5CA1 in B. Training phase: Control + ACSFCA1: n = 6; PolyI:C + ACSFCA1: n = 6; PolyI:C + AntiCA1: n = 6; PolyI:C + AntiCA3: n = 6; PolyI:C + Pep5CA1: n = 6. Memory test: Control + ACSFCA1: n = 6; PolyI:C + ACSFCA1: n = 4; PolyI:C + AntiCA1: n = 7; PolyI:C + AntiCA3: n = 7; PolyI:C + Pep5CA1: n = 7.

Article Snippet: The cleavage-resistant proBDNF (2 ng/ml; Cat#B257 Alomone Labs), TAT-Pep5 (4 ng/μl; Cat#506181, EMD Millipore), or artificial CSF (ACSF) was infused bilaterally at a rate of 0.5 μl/min/side for 2 min.

Techniques: Blocking Assay, Activation Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Maternal immune activation-induced proBDNF-mediated neural information processing dysfunction at hippocampal CA3-CA1 synapses associated with memory deficits in offspring

doi: 10.3389/fcell.2022.1018586

Figure Lengend Snippet: (A) The changes in EPSC of prymidal CA1 neurons. Typical consecutive sample traces of sEPSCs from each group (Top). The frequency of sEPSC (B) is not altered but the amplitude of sEPSC (C) is decreased in polyI:C-treated offspring. Incubation with anti-proBDNF antibody or TAT-Pep5 inhibitor can significantly enhance the declined amplitude. Data are presented as mean ± SEM. * p < 0.05, vs. Control + ACSFCA1, PolyI:C + AntiCA1 and PolyI:C + Pep5CA1. Control + ACSFCA1: n = 5; PolyI:C + ACSFCA1: n = 5; PolyI:C + AntiCA1: n = 7; PolyI:C + AntiCA3: n = 7; PolyI:C + Pep5CA1: n = 6.

Article Snippet: The cleavage-resistant proBDNF (2 ng/ml; Cat#B257 Alomone Labs), TAT-Pep5 (4 ng/μl; Cat#506181, EMD Millipore), or artificial CSF (ACSF) was infused bilaterally at a rate of 0.5 μl/min/side for 2 min.

Techniques: Incubation

Journal: Journal of Neuroinflammation

Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4 + T cells

doi: 10.1186/s12974-020-01850-0

Figure Lengend Snippet: Exogenous proBDNF protein reduced CD4 + T cells but increased CD8 + T cells of septic mice in vitro. Mice injected with saline or LPS (5 mg kg −1 ) for 5 days and the splenocytes were isolated and cultured for 3 days in vitro. Exogenous proBDNF did not alter the percentage of a CD3 + T cells in CD45 + cells or the percentage of b CD4 + T cells or c CD8 + T cells in CD3 + T cells in splenocytes from mice treated with saline. d Exogenous proBDNF did not alter the percentage of CD3 + T cells in CD45 + cells in splenocytes in septic mice. e–f ProBDNF treatment significantly decreased the percentage of e CD4 + T cells but increased the percentage of f CD8 + T cells in CD3 + T cells in splenocytes in LPS-treated mice. n = 4 in each group. Data were analyzed by one-way ANOVA and followed by Tukey post hoc test, * P < 0.05, ** P < 0.01. Data are presented as mean ± SEM. NT = no treated control. proB = exogenous proBDNF protein treatment

Article Snippet: 4 × 10 5 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml −1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

Techniques: In Vitro, Injection, Isolation, Cell Culture