Journal: Scientific Reports

Article Title: A comparison of commercial assays quantifying mature brain-derived neurotrophic factor (mBDNF) and its precursor (pro-BDNF) in human serum

doi: 10.1038/s41598-025-22278-7

Figure Lengend Snippet: Summary of ELISA’s characteristics. Main characteristics declared by the manufacturers of each ELISA kit evaluated in this study.

Article Snippet: Samples were prepared with 16 ng of human - pro-BDNF (Cat. #B-256-B) from Alomone Labs (Jerusalem, Israel), mature BDNF (Cat. #orb1675341), from Biorbyt (Cambridge, UK), and Bovine Serum Albumin (BSA) as negative control (Cat. #A7030, Sigma-Aldrich, St. Louis, MO) mixed with 4X Laemmli sample buffer (Cat. #1610747, Bio-Rad Laboratories, CA, USA) containing 10% β-mercaptoethanol (Cat. #M3148, Sigma-Aldrich, St. Louis, MO) and milliQ-water.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: A comparison of commercial assays quantifying mature brain-derived neurotrophic factor (mBDNF) and its precursor (pro-BDNF) in human serum

doi: 10.1038/s41598-025-22278-7

Figure Lengend Snippet: ELISA kits tested. Comparison between the performance declared by the manufacturer and the one obtained in this study for each ELISA kit tested.

Article Snippet: Samples were prepared with 16 ng of human - pro-BDNF (Cat. #B-256-B) from Alomone Labs (Jerusalem, Israel), mature BDNF (Cat. #orb1675341), from Biorbyt (Cambridge, UK), and Bovine Serum Albumin (BSA) as negative control (Cat. #A7030, Sigma-Aldrich, St. Louis, MO) mixed with 4X Laemmli sample buffer (Cat. #1610747, Bio-Rad Laboratories, CA, USA) containing 10% β-mercaptoethanol (Cat. #M3148, Sigma-Aldrich, St. Louis, MO) and milliQ-water.

Techniques: Enzyme-linked Immunosorbent Assay, Comparison

Journal: Scientific Reports

Article Title: A comparison of commercial assays quantifying mature brain-derived neurotrophic factor (mBDNF) and its precursor (pro-BDNF) in human serum

doi: 10.1038/s41598-025-22278-7

Figure Lengend Snippet: Levels of proteins (ng/mL) obtained in human serum samples ( n = 23). Different dilutions were assessed in ELISA kits from R&D Systems ( A , B and E ) [total BDNF Quantikine ELISA kit#DBNT00 (A) , Human pro-BDNF DuoSet ELISA kit #DY3175 (B) and human free BDNF Quantikine ELISA kit #DBD00 (E) ], Aviscera Bioscience ( C and F ) [high sensitivity pro-BDNF human ELISA kit #SK00752-09 (C) and high sensitive BDNF ELISA kit #SK00752-01 (F) ] and Finetest ( D and G ) [human pro-BDNF ELISA kit #EH4255 (D) and human BDNF ELISA kit #EH0043 (G) ]. Comparison of protein levels obtained in human serum samples diluted at 1:20 (H) for pro-BDNF and at 1:40 (I) for BDNF (common dilutions) in the ELISA kits from the 3 companies used in this study (R&D Systems, Aviscera Bioscience and FineTest). Each value obtained of the samples analysed was represented, and the median ± SD of all of them.

Article Snippet: Samples were prepared with 16 ng of human - pro-BDNF (Cat. #B-256-B) from Alomone Labs (Jerusalem, Israel), mature BDNF (Cat. #orb1675341), from Biorbyt (Cambridge, UK), and Bovine Serum Albumin (BSA) as negative control (Cat. #A7030, Sigma-Aldrich, St. Louis, MO) mixed with 4X Laemmli sample buffer (Cat. #1610747, Bio-Rad Laboratories, CA, USA) containing 10% β-mercaptoethanol (Cat. #M3148, Sigma-Aldrich, St. Louis, MO) and milliQ-water.

Techniques: Blood/Plasma/Serum Samples, Enzyme-linked Immunosorbent Assay, Sensitivity, Comparison

Journal: Scientific Reports

Article Title: A comparison of commercial assays quantifying mature brain-derived neurotrophic factor (mBDNF) and its precursor (pro-BDNF) in human serum

doi: 10.1038/s41598-025-22278-7

Figure Lengend Snippet: Western blot for qualitative analysis of pro-BDNF and mBDNF antibodies. Antibodies from R&D Systems [total BDNF Quantikine #DBNT00 (A) , human pro-BDNF DuoSet #DY3175 (B) and human free BDNF Quantikine #DBD00 (C) ], FineTest [human pro-BDNF #EH4255 (D) and human BDNF #EH0043 (E) ], and Aviscera Bioscience [high sensitivity pro-BDNF human #SK00752-09 (F) and high sensitive BDNF #SK00752-01 (G) ] ELISA kit were tested. Original blots are presented in Supplementary Figure S6.

Article Snippet: Samples were prepared with 16 ng of human - pro-BDNF (Cat. #B-256-B) from Alomone Labs (Jerusalem, Israel), mature BDNF (Cat. #orb1675341), from Biorbyt (Cambridge, UK), and Bovine Serum Albumin (BSA) as negative control (Cat. #A7030, Sigma-Aldrich, St. Louis, MO) mixed with 4X Laemmli sample buffer (Cat. #1610747, Bio-Rad Laboratories, CA, USA) containing 10% β-mercaptoethanol (Cat. #M3148, Sigma-Aldrich, St. Louis, MO) and milliQ-water.

Techniques: Western Blot, Analysis, Sensitivity, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: A comparison of commercial assays quantifying mature brain-derived neurotrophic factor (mBDNF) and its precursor (pro-BDNF) in human serum

doi: 10.1038/s41598-025-22278-7

Figure Lengend Snippet: Correlation’s heatmaps. Results about the Spearman correlations obtained among serum protein levels (total, pro-BDNF and BDNF) obtained in the different commercial brands used in this study [R&D Systems, Aviscera Bioscience and FineTest].* p-value < 0.05.

Article Snippet: Samples were prepared with 16 ng of human - pro-BDNF (Cat. #B-256-B) from Alomone Labs (Jerusalem, Israel), mature BDNF (Cat. #orb1675341), from Biorbyt (Cambridge, UK), and Bovine Serum Albumin (BSA) as negative control (Cat. #A7030, Sigma-Aldrich, St. Louis, MO) mixed with 4X Laemmli sample buffer (Cat. #1610747, Bio-Rad Laboratories, CA, USA) containing 10% β-mercaptoethanol (Cat. #M3148, Sigma-Aldrich, St. Louis, MO) and milliQ-water.

Techniques:

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, Confocal 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: Scientific reports

Article Title: Cognitive and sensorimotor benefits of moderate- and high-intensity exercise are associated with specific expression of neurotrophic markers in older rats.

doi: 10.1038/s41598-025-90719-4

Figure Lengend Snippet: Fig. 1. Experimental protocol. All trained animals carried out 4 weeks of training, 5 times per week either in MICT or HIIT. All animals performed both incremental test and grip strength at PRE, 2W and POST, while they only performed adhesive removal test and the novel object recognition test at the end of the training (in POST). During the incremental test, blood lactate was collected at every level to determine the SLT. At the end of the protocol (48 h after the last incremental test) both hippocampi and cortices were removed to perform Western blot and ELISA measurements.

Article Snippet: Quantification of pro and mature BDNF standard and cortical samples was respectively performed with proBDNF Rapid ELISA Kit (Biosensis®, BEK-2217-2P – sandwich ELISA–Thebarton, SA, Australia) and mBDNF Rapid ELISA Kit Kit (Biosensis®, BEK-2211-1P/2P–sandwich ELISA–Thebarton, SA, Australia) in the concentrated solutions following the manufacturer’s protocol.

Techniques: Adhesive, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Scientific reports

Article Title: Cognitive and sensorimotor benefits of moderate- and high-intensity exercise are associated with specific expression of neurotrophic markers in older rats.

doi: 10.1038/s41598-025-90719-4

Figure Lengend Snippet: Fig. 4. Effect of MICT and HIIT programs on cortical protein levels. (a) Level of cortical mBDNF (n = 19) measured by ELISA (in pg/ml-1) in MICT, HIIT and Control groups at POST training. Level of cortical (b) TrkB (n = 20), (c) pTrkB (n = 20), (d), p757NTR (n = 19), (e) NKCC1/KCC2 ratio (n = 19), (f) KCC2 (n = 18) and (g) CTSB (n = 20) in MICT, HIIT and Control groups at POST training. Protein level measured by Western blot, was calculated as the ratio of the protein of interest divided by the protein of normalization relative to the control. The normalization protein was the α-tubulin for all the molecules except for the KCC2 which was β3-tubulin. Pictures of Western blot membranes are shown below each graph. Dashed lines were added to separate the groups. Note that the α-tubulin used for TrkB (c) was the same used for pTrkB (d), as membranes were stripped between these two quantifications. The samples derived from the same experiment and that gels/ blots were systematically processed in parallel the same day (2 gels per protein to reach a sufficient number of samples). *Significant differences in protein levels between groups. Data is expressed in mean ± SD (a–f) and in median (min to max) (g).

Article Snippet: Quantification of pro and mature BDNF standard and cortical samples was respectively performed with proBDNF Rapid ELISA Kit (Biosensis®, BEK-2217-2P – sandwich ELISA–Thebarton, SA, Australia) and mBDNF Rapid ELISA Kit Kit (Biosensis®, BEK-2211-1P/2P–sandwich ELISA–Thebarton, SA, Australia) in the concentrated solutions following the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Derivative Assay

Journal: Scientific reports

Article Title: Cognitive and sensorimotor benefits of moderate- and high-intensity exercise are associated with specific expression of neurotrophic markers in older rats.

doi: 10.1038/s41598-025-90719-4

Figure Lengend Snippet: Fig. 5. Effect of MICT and HIIT programs on hippocampal protein levels. (a) Level of hippocampal IGF- 1(n = 21) measured by ELISA in MICT, HIIT and Control groups at POST training. Level of hippocampal (b) IGFB-2 (n = 21), (c) p75NTR (n = 22), (d) TrkB (n = 21), (e) CTSB (n = 22), (f) NKCC1 (n = 21), (g) KCC2 (n = 20), and (h) the NKCC1/KCC2 ratio (n = 18) measured by Western blot in MICT, HIIT and Control groups at POST. Protein level was calculated as the ratio of the protein of interest divided by the protein of normalization relative to the control. The normalization protein was the α-tubulin for all the molecules except for the KCC2 which was β3-tubulin. Pictures of the Western blot membranes are shown below each graph. Dashed lines were added to separate the groups. Note that the α-tubulin used for p75NTR (c) was the same used for NKCC1 (f) as their molecular weight are different, the quantification of both was conducted on the same membranes. The samples derived from the same experiment and that gels/blots were processed in parallel the same day (2 gels per protein to reach a sufficient number of samples). *Significant differences in protein levels between groups. All data is expressed in mean ± SD.

Article Snippet: Quantification of pro and mature BDNF standard and cortical samples was respectively performed with proBDNF Rapid ELISA Kit (Biosensis®, BEK-2217-2P – sandwich ELISA–Thebarton, SA, Australia) and mBDNF Rapid ELISA Kit Kit (Biosensis®, BEK-2211-1P/2P–sandwich ELISA–Thebarton, SA, Australia) in the concentrated solutions following the manufacturer’s protocol.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Western Blot, Molecular Weight, Derivative Assay