Human TrkB Antibody

Detection of TrkB in Baf3 cells by Flow Cytometry. Baf3 cells transfected with Human TrkB (filled histogram) vs Irrelevant transfectant cells (open histogram) were stained with Mouse Anti-Human TrkB Monoclonal Antibody (Catalog # MAB3971) followed by Fluorescein-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0103B). View our protocol for Staining Membrane-associated Proteins.

Detection of TrkB by Flow Cytometry DLBCL cell lines express neurotrophin receptors.A: RT-PCR detection of p75NTR, and its co-receptor sortilin, TrkA and truncated gp95TrkB mRNA was performed on SUDHL cells cultured with 10% FCS for 72 h. beta -actin was included as a control of cDNA quality. B: The truncated TrkB, p75NTR and sortilin receptor protein expression was confirmed by western blotting of cell lysates. Blots were reprobed with anti- beta -actin as a loading control. C: Flow cytometry analysis demonstrating TrkB membrane detection expressed in percentage of positive cells (bold line) in unpermeabilized SUDHL6 cells in contrast to SUDHL4 cells, whereas both cell lines expressed intracellular TrkB (dotted line: the isotypic controls). D: Immunofluorescence staining was performed, as described in Materials and Methods, in DLBCL cell lines that confirmed surface TrkB expression (in green) in some BDNF (inset in red) positive SUDHL6 cells (in blue: DAPI staining). The neuroblastoma cell line, IMR32, was used as a positive control. Data are representative of four independent experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/22076137), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of TrkB by Flow Cytometry Modulation of neurotrophin and TrkB expression by SHDHL4 cells is induced by serum deprivation.A: RT-PCR analysis of NGF, BDNF, TrkA, gp95TrkB, p75NTR and sortilin mRNA on SUDHL4 cells submitted to 72 h serum deprivation (−FCS) as compared to standard culture conditions (10% FCS, +FCS). beta -actin was included as a control of cDNA quality. B: Western blott detection of truncated TrkB and p75NTR protein expression. Blots were reprobed with anti- beta -actin as a loading control. C: Immunoprecipitation of sortilin (IP) and western blotting analysis (IB, immunoblotting with anti-p75NTR or anti-sortilin as control) demonstrating the association of sortilin with p75NTR in cell lysates of both culture conditions. D: secretion of BDNF detected in cell supernatants after immunoprecipitation and western blotting. E: Flow cytometry analysis demonstrating membranous expression of TrkB (bold line) and intracellular TrkB, NGF (bold line) and BDNF expressions expressed in percentage of SUDHL4 positive cells cultured with or without FCS (dotted line: isotypic controls). Data are representative of four independent experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/22076137), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of TrkB by Western Blot TrkB expression in human brain and platelets. (A) Healthy human cortex (C; 3 µg) and platelets lysates (platelets; 30 to 75 µg) were analyzed in denaturing and reducing conditions and blotted with antibodies raised against TrkB extracellular domain. Left: molecular weight marker in kDa. Right: antibody catalog number. beta -actin was used as a loading control. (B) Human glioblastoma cells U87-MG and healthy platelets lysates were either left untreated (Untreated) or submitted to 37°C overnight in absence (Sham) or presence of N-deglycosylation enzyme PNGase F (PNGase F). Membranes were blotted with ANT-019 antibody against TrkB ECD. PNGase F activity was confirmed by reblotting membranes with antibodies against N-glycosylated proteins sortilin for U87-MG and CD42b for platelets. (C) Human glioblastoma U87-MG cells and healthy human platelets isolated from whole blood were fixed or fixed and permeabilized. Cells were labeled with antibodies directed toward the extracellular domain of the TrkB receptor and analyzed by flow cytometry. Results shown are representative of (A) ≥ 3 independent experiments and ≥ 5 different platelet samples from different donors and (B, C) 3 independent experiments and 3 different donors for platelets samples. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33643311), licensed under a CC-BY license. Not internally tested by R&D Systems.