TrkB Antibody (72509) [Unconjugated]

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.

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 (//pubmed.ncbi.nlm.nih.gov/22076137), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/22076137), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/33643311), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: Flow, CyTOF-ready
• Clone: 72509
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

TrkB Antibody (72509) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human TrkB
  Cys32-His430
  Accession # Q16620
• Specificity: Detects human TrkB in direct ELISAs and Western blots. In Western blots, no cross-reactivity with recombinant human (rh) TrkA, recombinant rat TrkA, recombinant mouse (rm) TrkB, rhTrkC, or rmTrkC is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: NTRK2
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 0.25 ug/10^6 cells

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for TrkB Antibody (72509) [Unconjugated]

• BDNF/NT-3 growth factors receptor
• EC 2.7.10
• EC 2.7.10.1
• GP145-TrkB
• Neurotrophic tyrosine kinase receptor type 2
• neurotrophic tyrosine kinase, receptor, type 2
• NTRK2
• TrkB tyrosine kinase
• TrkB
• trk-B
• TRKBBDNF-tropomyosine receptor kinase B
• tyrosine kinase receptor B

Background

The neurotrophins, including NGF, BDNF, NT-3, and NT-4/5 constitute a group of structurally related, secreted proteins that play an important role in the development and function of the nervous system. The biological activities of the neurotrophins are mediated by binding to the different members of the Trk family tyrosine kinase receptors. Three Trk family proteins, TrkA, TrkB, and TrkC, exhibiting different ligand specificities, have been identified. TrkA binds NGF, TrkB binds BDNF and
NT‑4/5 and TrkC binds NT-3. All Trk family proteins share a conserved complex subdomain organization consisting of a signal peptide, two cysteine-rich domains, a cluster of three leucine-rich motifs, and two immunoglobulin-like domains in the extracellular region, as well as an intracellular region that contains the tyrosine kinase domain. Natural splice variants of the different Trks, including TrkB variants lacking the first cysteine-rich domain, the first and second or all three of the leucine-rich motifs, or the tyrosine kinase domain, have been described. The role of the different extracellular subdomains of TrkB in mediating neurotrophin binding and discrimination is currently being investigated. At the protein sequence level, human and rat TrkB are greater than 90% identical and the proteins exhibit cross-species activity. TrkB is primarily expressed in the nervous system. However, low levels of TrkB expression have also been observed in a wide variety of tissues (pancreas, kidneys, ovary) outside the nervous system.

1. Ninkina, N. et al. (1997) J. Biol. Chem. 272:13019.
2. Middlemas, D.S. et al. (1991) Mol. Cell Biol. 11:143.
3. Soppet, D. et al. (1991) Cell 65:895.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for TrkB Antibody (MAB3971)(12)

Publications using MAB3971

• Hiranuma, M;Okuda, Y;Fujii, Y;Richard, JP;Watanabe, T; Characterization of human iPSC-derived sensory neurons and their functional assessment using multi electrode array Scientific reports 2024-03-12 [PMID: 38472288] (Immunocytochemistry, Human)
• Kenyi Saito-Diaz, Christina James, Archie Jayesh Patel, Nadja Zeltner Isolation of human pluripotent stem cell-derived sensory neuron subtypes by immunopanning Frontiers in Cell and Developmental Biology 5/3/2023 [PMID: 37206924]
• Kenyi Saito-Diaz, Nadja Zeltner A protocol to differentiate nociceptors, mechanoreceptors, and proprioceptors from human pluripotent stem cells STAR Protocols 6/1/2022 [PMID: 35330962]
• Fleury S, Boukhatem I, Le Blanc J et al. Tissue-Specificity of Antibodies Raised Against TrkB and p75NTR Receptors; Implications for Platelets as Models of Neurodegenerative Diseases Frontiers in Immunology 2/11/2021 [PMID: 33643311]
• T Guerenne-D, V Couderc, L Duponchel, V Sol, P Leproux, JM Petit Multiplex coherent anti-Stokes Raman scattering microspectroscopy detection of lipid droplets in cancer cells expressing TrkB Sci Rep, 2020-10-07;10(1):16749. 2020-10-07 [PMID: 33028922] (Western Blot, Human)
• A Abbaci, H Talbot, S Saada, N Gachard, J Abraham, A Jaccard et al. Neurotensin receptor type 2 protects B-cell chronic lymphocytic leukemia cells from apoptosis Oncogene 2/1/2018 [PMID: 29059151]
• Stefanie Traub, Heiko Stahl, Holger Rosenbrock, Eric Simon, Lore Florin, Lisa Hospach et al. Pharmaceutical Characterization of Tropomyosin Receptor Kinase B-Agonistic Antibodies on Human Induced Pluripotent Stem (hiPS) Cell–Derived Neurons Journal of Pharmacology and Experimental Therapeutics 6/1/2017 [PMID: 28351853]
• Stefanie Traub, Heiko Stahl, Holger Rosenbrock, Eric Simon, Lore Florin, Lisa Hospach et al. Pharmaceutical Characterization of Tropomyosin Receptor Kinase B-Agonistic Antibodies on Human Induced Pluripotent Stem (hiPS) Cell–Derived Neurons Journal of Pharmacology and Experimental Therapeutics 2017-06-01 [PMID: 28351853] (ELISA Detection, Human)
• Michael J. Shamblott, Marci L. O’Driscoll, Danielle L. Gomez, Dustin L. McGuire Neurogenin 3 is regulated by neurotrophic tyrosine kinase receptor type 2 (TRKB) signaling in the adult human exocrine pancreas Cell Communication and Signaling 12/1/2016 [PMID: 27659207]
• Giraud S, Loum E, Bessette B, Mathonnet M, Lalloue F P75 neurotrophin receptor is sequestered in the Golgi apparatus of the U-87 MG human glioblastoma cell line. Int. J. Oncol., 2010-12-06;38(2):391-9. 2010-12-06 [PMID: 21152857] (ICC, Human)
• Berzi A, Ayata CK, Cavalcante P, Falcone C, Candiago E, Motta T, Bernasconi P, Hohlfeld R, Mantegazza R, Meinl E, Farina C BDNF and its receptors in human myasthenic thymus: implications for cell fate in thymic pathology. J. Neuroimmunol., 2008-06-13;197(2):128-39. 2008-06-13 [PMID: 18555538] (Flow Cytometry, ICC, IHC-Fr, Human)
• Kajiya M, Shiba H, Fujita T, Ouhara K, Takeda K, Mizuno N, Kawaguchi H, Kitagawa M, Takata T, Tsuji K, Kurihara H Brain-derived neurotrophic factor stimulates bone/cementum-related protein gene expression in cementoblasts. J. Biol. Chem., 2008-04-03;283(23):16259-67. 2008-04-03 [PMID: 18390540] (ICC, Human)

Bioinformatics

• Gene Symbol: NTRK2
• Uniprot:
  • Q16620()