anti probdnf antibody  (Alomone Labs)

Journal: Frontiers in Pharmacology

Article Title: Pentylenetetrazole kindling impairs place recognition memory associated with suppressing proBDNF-mediated neural information flows at the hippocampal CA3-CA1 synapses

doi: 10.3389/fphar.2026.1881525

Figure Lengend Snippet: The changes in neuronal firing rates and EPSCs of CA1 putative FS interneurons. The firing rates of both RS (A) and putative FS (B) cells were significantly increased following behavioral training in the control animals while only the frequency of putative FS was abnormally increased after the training. Representative firing traces of RS pyramidal neurons and putative FS interneurons are shown as upper insets in panels (A,B) , respectively. #, P < 0.05, vs. control; *, P < 0.05, vs. control and control-L; %, P < 0.05, vs. epilepsy-L. (C) Representative traces of sEPSCs recorded from putative FS interneurons. A magnified segment of each trace is shown to clearly present individual sEPSC waveform and peak amplitude. Scale bars for sEPSCs: 0.5 s (horizontal) and 5 pA (vertical). Both frequency (D) and amplitude (E) of sEPSC were drastically enhanced following behavioral training in PTZ-treated epileptic animals. Incubation with anti-proBDNF antibody or TAT-Pep5 inhibitor could significantly suppressed the enhanced frequency and amplitude. (F) Representative traces of mEPSCs recorded from putative FS interneurons in the presence of TTX. Scale bars for mEPSCs: 0.5 s (horizontal) and 5 pA (vertical). (G) Mean frequency and (H) amplitude of mEPSCs. Both parameters were increased in the Epilepsy-L group and rescued by anti-proBDNF antibody or TAT-Pep5. *, P < 0.05, vs. others. The number of rats in each group was indicated in figure legend.

Article Snippet: Either anti-proBDNF antibody (10 μg/μL; Cat#ANT-006, Alomone Labs), TAT-Pep5 (4 ng/μL; Cat#506181, EMD Millipore), K252a (25 μg/μL; Cat#82497; Sigma-Aldrich) or artificial CSF (ACSF, Cat#3525, Tocris Bioscience) was infused at a rate of 0.5 μL/min/side for 2 min.

Techniques: Control, Incubation, Patch Clamp

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Long non-coding SINEUP RNA enhancing BDNF translation prevents dendritic atrophy following an Aβ-peptide challenge

doi: 10.1007/s00018-026-06175-4

Figure Lengend Snippet: SINEUP strategy, specificity and efficacy in targeting BDNF protein upregulation. A-B . Rat BDNF gene structure with schematic representation of BDNF transcripts characterised by multiple alternative splicing of 5’ and 3’UTRs (grey boxes), a coding sequence (CDS, white boxes) and two poly-adenylation sites (polyA). B. SINEUP-PAN and SINEUP-Ex1 annealing sites on BDNF transcripts used in this study. C-H. HEK293T cells are co-transfected with different plasmids to demonstrate the efficacy of SINEUP-PAN or SINEUP-Ex1 on the indicated BDNF transcripts with respect to control (GFP). Plasmids were transfected in 1:3 ratio. Tubulin (55 kDa) is indicative of gel loading and is used for normalization in the Western blot analysis. Histograms quantify average data from n = 4 experiments. The molecular weight for the BDNF-GFP band of 64–65 KDa correspond to the predicted size for a chimeric protein made of the proBDNF form (32–34 KDa) and the GFP (27–30 KDa). C . Effect of SINEUP-PAN on protein levels encoded by the transfected chimeric Ex1-BDNF-GFP transcript with respect to control represented by the plasmid encoding the reporter protein GFP alone. p = 0.0005. D . Effect of SINEUP-Ex1 on protein levels encoded by the transfected chimeric Ex1-BDNF-GFP transcript. p = 0.0299. E-F . Effect of SINEUP-PAN on protein levels encoded by the transfected chimeric Ex2B-BDNF-GFP (E) and chimeric Ex2C-BDNF-GFP (F) transcripts. n = 3. G-H . Effect of SINEUP-Ex1 on chimeric Ex2B-BDNF-GFP (G) and chimeric Ex2C-BDNF-GFP (H) transcripts with respect to control (GFP). All the quantitative results shown are produced from a repetition of n = 3 experiments and the Western blot images show the greatest effect obtained and therefore, are not representative of the average results

Article Snippet: The membrane was blocked by 5% non-fat milk in TBST- solution (1 h, RT) and then, the HEK293T cells were incubated with the rabbit polyclonal anti proBDNF antibody (Alomone, Cat. # ANT-006, dilution 1:1000, over-night at 4 °C) while the SH-SY5Y cells were incubated also with the mouse monoclonal anti-BDNF recognizing both BDNF forms (Sigma Cat. #B5050, dilution 1:1000, over-night at 4 °C). and mouse anti alpha tubulin (Sigma, Cat. # T6074, dilution 1:3000) over-night at 4 °C.

Techniques: Alternative Splicing, Sequencing, Transfection, Control, Western Blot, Molecular Weight, Plasmid Preparation, Produced

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Long non-coding SINEUP RNA enhancing BDNF translation prevents dendritic atrophy following an Aβ-peptide challenge

doi: 10.1007/s00018-026-06175-4

Figure Lengend Snippet: SINEUP strategy, specificity and efficacy in targeting BDNF protein upregulation. A-B . Rat BDNF gene structure with schematic representation of BDNF transcripts characterised by multiple alternative splicing of 5’ and 3’UTRs (grey boxes), a coding sequence (CDS, white boxes) and two poly-adenylation sites (polyA). B. SINEUP-PAN and SINEUP-Ex1 annealing sites on BDNF transcripts used in this study. C-H. HEK293T cells are co-transfected with different plasmids to demonstrate the efficacy of SINEUP-PAN or SINEUP-Ex1 on the indicated BDNF transcripts with respect to control (GFP). Plasmids were transfected in 1:3 ratio. Tubulin (55 kDa) is indicative of gel loading and is used for normalization in the Western blot analysis. Histograms quantify average data from n = 4 experiments. The molecular weight for the BDNF-GFP band of 64–65 KDa correspond to the predicted size for a chimeric protein made of the proBDNF form (32–34 KDa) and the GFP (27–30 KDa). C . Effect of SINEUP-PAN on protein levels encoded by the transfected chimeric Ex1-BDNF-GFP transcript with respect to control represented by the plasmid encoding the reporter protein GFP alone. p = 0.0005. D . Effect of SINEUP-Ex1 on protein levels encoded by the transfected chimeric Ex1-BDNF-GFP transcript. p = 0.0299. E-F . Effect of SINEUP-PAN on protein levels encoded by the transfected chimeric Ex2B-BDNF-GFP (E) and chimeric Ex2C-BDNF-GFP (F) transcripts. n = 3. G-H . Effect of SINEUP-Ex1 on chimeric Ex2B-BDNF-GFP (G) and chimeric Ex2C-BDNF-GFP (H) transcripts with respect to control (GFP). All the quantitative results shown are produced from a repetition of n = 3 experiments and the Western blot images show the greatest effect obtained and therefore, are not representative of the average results

Article Snippet: The membrane was blocked by 5% non-fat milk in TBST- solution (1 h, RT) and then, the HEK293T cells were incubated with the rabbit polyclonal anti proBDNF antibody (Alomone, Cat. # ANT-006, dilution 1:1000, over-night at 4 °C) while the SH-SY5Y cells were incubated also with the mouse monoclonal anti-BDNF recognizing both BDNF forms (Sigma Cat. #B5050, dilution 1:1000, over-night at 4 °C).

Techniques: Alternative Splicing, Sequencing, Transfection, Control, Western Blot, Analysis, Molecular Weight, Plasmid Preparation, Produced