TrkB binds to BDNF at the plasma membrane.Live TIRF imaging of TrkB-ACP-488 and labeled BDNF co-movement in HEK293T (HEK). Cells treated with streptavidin-647-labeled human BDNF-Biotin (#B-250-B) for 5 minutes and imaged at 8 fps. Arrowhead marks co-localized TrkB-BDNF particle. Scale bar = 2 µm. Right: color trace shows movement trajectory of TrkB-ACP-488 and BDNF-647 particles over time (blue- start, red- end of trajectory) Scale bar = 1 µm.Unpublished data from Zahavi, E.E.et al. (2018) Sci. Signal.11, eaao4006. Figure generously provided by Dr. Eran Perlson, Dept. of Physiology and Pharmacology, Tel-Aviv University, Tel-Aviv, Israel.
BDNF co-localizes with TrkB in mouse cerebellum region.
A. Fixed brain sections were incubated with human BDNF-Biotin (#B-250-B) followed by Streptavidin-Cy3. B. Same sections were incubated with Anti-TrkB (extracellular)-ATTO Fluor-488 Antibody (#ANT-019-AG). C. Merge of A and B reveals co-localization in the Purkinje layer (P). TrkB positive cells (green) with BDNF binding (red, arrow) is seen.
Alomone Labs human BDNF-Biotin induces ERK1/2 MAPK phosphorylation in transfected HEK-293 cells as BDNF.
Transfected cells were serum depleted for 2 h, 3 days post-transfection and then challenged with 1 and 10 ng/ml Recombinant human BDNF protein (#B-250) or 1, 10 and 100 ng/ml human BDNF-Biotin (#B-250-B) for 10 min. The cell proteins were resolved by SDS-PAGE and detected with anti-phospho-ERK1/2.
Alomone Labs human BDNF-Biotin induces ERK1/2 MAPK phosphorylation in mouse cortical neurons as BDNF.
Mouse cortical neurons were starved for 4 h and then challenged with 50 ng/ml Recombinant human BDNF protein (#B-250) or human BDNF-Biotin (#B-250-B) for 30 min. 20 µg of the cell protein lysate were resolved by SDS-PAGE and detected with anti-phospho-ERK1/2 and anti-tubulin (the experiment was performed and the picture was kindly provided by Dr. Eran Perlsson, the Dept. of Pharmacology and Physiology, Tel-Aviv University).
Alomone Labs human BDNF-Biotin binds TrKB receptors in living and stably transfected HEK293 cells and is displaced by un-labeled human BDNF.
Living cells were incubated with 100 ng/ml human BDNF-Biotin (#B-250-B) in the absence (A-D) and presence (E,F) of 2 µg/ml Recombinant human BDNF protein (#B-250). Pictures were taken in early stages of the experiment (A-B) as well as in late stages (C-D) in order to show the accumulation effect of human BDNF-Biotin. human BDNF-Biotin was visualized using StrepAvidin-AlexaFluor-594. Similar results were observed using 10 ng/ml human BDNF-Biotin (data not shown).
BDNF co-localizes with TrkB in mouse hippocampal CA1 region.
A. Fixed brain sections were incubated with human BDNF-Biotin (#B-250-B) followed by Streptavidin-Cy3. B. Same sections were incubated with Anti-TrkB (extracellular)-ATTO Fluor-488 Antibody (#ANT-019-AG). C. Merge of A and B reveals co-localization in the pyramidal layer (P). Outside the pyramidal layer, there are also cells with only BDNF (red, horizontal arrows) or only TrkB expression (green, vertical arrows).
