Overview

  • Product name

    Anti-TrkA antibody [BS470] - Extracellular domain
    See all TrkA primary antibodies
  • Description

    Mouse monoclonal [BS470] to TrkA - Extracellular domain
  • Host species

    Mouse
  • Specificity

    Does not cross-react with TrkB or TrkC.

  • Tested applications

    Suitable for: ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human TrkA aa 235-246 (extracellular).
    Sequence:

    (c)SATVMKSGGLPS


    Database link: P04629

  • Positive control

    • Flow Cyt: SH-SY5Y and DU 145 cells. ICC/IF: SH-SY5Y cells.

Properties

  • Form

    Lyophilised:Reconstitute in 100 uL of sterile water. Centrifuge to remove any insoluble material. Final buffer contains no preservatives. Highest purity glycerol (1:1) may be added for additional stability.
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 8.00
    Constituents: Phosphate Buffer, 1.74% Sodium chloride, 3% Trehalose
  • Concentration information loading...
  • Purity

    Protein G purified
  • Purification notes

    Purified from TCS.
  • Clonality

    Monoclonal
  • Clone number

    BS470
  • Isotype

    IgG3
  • Light chain type

    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab244209 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 1 - 5 µg/ml.

Flow cytometry data and immunofluorescence staining of TrkA (extracellular domain) receptor in SHSY-5Y cells has shown that detection of TrkA expression depends on its cellular localization (membrane vs. internal stores). Thus, permeabilization of cells may be required, despite that clone BS470 was raised against the extracellular domain of TrkA.

Flow Cyt Use a concentration of 20 µg/ml.

Flow cytometry data and immunofluorescence staining of TrkA (extracellular domain) receptor in SHSY-5Y cells has shown that detection of TrkA expression depends on its cellular localization (membrane vs. internal stores). Thus, permeabilization of cells may be required, despite that clone BS470 was raised against the extracellular domain of TrkA.

Target

  • Function

    Receptor tyrosine kinase involved in the development and the maturation of the central and peripheral nervous systems through regulation of proliferation, differentiation and survival of sympathetic and nervous neurons. High affinity receptor for NGF which is its primary ligand, it can also bind and be activated by NTF3/neurotrophin-3. However, NTF3 only supports axonal extension through NTRK1 but has no effect on neuron survival. Upon dimeric NGF ligand-binding, undergoes homodimerization, autophosphorylation and activation. Recruits, phosphorylates and/or activates several downstream effectors including SHC1, FRS2, SH2B1, SH2B2 and PLCG1 that regulate distinct overlapping signaling cascades driving cell survival and differentiation. Through SHC1 and FRS2 activates a GRB2-Ras-MAPK cascade that regulates cell differentiation and survival. Through PLCG1 controls NF-Kappa-B activation and the transcription of genes involved in cell survival. Through SHC1 and SH2B1 controls a Ras-PI3 kinase-AKT1 signaling cascade that is also regulating survival. In absence of ligand and activation, may promote cell death, making the survival of neurons dependent on trophic factors.
    Isoform TrkA-III is resistant to NGF, constitutively activates AKT1 and NF-kappa-B and is unable to activate the Ras-MAPK signaling cascade. Antagonizes the anti-proliferative NGF-NTRK1 signaling that promotes neuronal precursors differentiation. Isoform TrkA-III promotes angiogenesis and has oncogenic activity when overexpressed.
  • Tissue specificity

    Isoform TrkA-I is found in most non-neuronal tissues. Isoform TrkA-II is primarily expressed in neuronal cells. TrkA-III is specifically expressed by pluripotent neural stem and neural crest progenitors.
  • Involvement in disease

    Congenital insensitivity to pain with anhidrosis
    Chromosomal aberrations involving NTRK1 are found in papillary thyroid carcinomas (PTCs) (PubMed:2869410, PubMed:7565764, PubMed:1532241). Translocation t(1;3)(q21;q11) with TFG generates the TRKT3 (TRK-T3) transcript by fusing TFG to the 3'-end of NTRK1 (PubMed:7565764). A rearrangement with TPM3 generates the TRK transcript by fusing TPM3 to the 3'-end of NTRK1 (PubMed:2869410). An intrachromosomal rearrangement that links the protein kinase domain of NTRK1 to the 5'-end of the TPR gene forms the fusion protein TRK-T1. TRK-T1 is a 55 kDa protein reacting with antibodies against the C-terminus of the NTRK1 protein (PubMed:1532241).
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. Insulin receptor subfamily.
    Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 2 LRR (leucine-rich) repeats.
    Contains 1 LRRCT domain.
    Contains 1 protein kinase domain.
  • Domain

    The transmembrane domain mediates interaction with KIDINS220.
    The extracellular domain mediates interaction with NGFR.
  • Post-translational
    modifications

    Ligand-mediated autophosphorylation. Interaction with SQSTM1 is phosphotyrosine-dependent. Autophosphorylation at Tyr-496 mediates interaction and phosphorylation of SHC1.
    N-glycosylated (Probable). Isoform TrkA-I is N-glycosylated.
    Ubiquitinated. Undergoes polyubiquitination upon activation; regulated by NGFR. Ubiquitination regulates the internalization of the receptor.
  • Cellular localization

    Cell membrane. Early endosome membrane. Late endosome membrane. Internalized to endosomes upon binding of NGF or NTF3 and further transported to the cell body via a retrograde axonal transport. Localized at cell membrane and early endosomes before nerve growth factor (NGF) stimulation. Recruited to late endosomes after NGF stimulation. Colocalized with RAPGEF2 at late endosomes (By similarity).
  • Information by UniProt
  • Database links

  • Alternative names

    • gp140trk antibody
    • High affinity nerve growth factor receptor antibody
    • High affinity nerve growth factor receptor precursor antibody
    • MTC antibody
    • Neurotrophic tyrosine kinase receptor type 1 antibody
    • NTRK1 antibody
    • NTRK1_HUMAN antibody
    • Oncogene TRK antibody
    • p14-TrkA antibody
    • p140 TrkA antibody
    • p140-TrkA antibody
    • Slow nerve growth antibody
    • Trk A antibody
    • TRK antibody
    • Trk-A antibody
    • TRK1 antibody
    • TRK1-transforming tyrosine kinase protein antibody
    • Tropomyosin-related kinase A antibody
    • Tyrosine kinase receptor A antibody
    • Tyrosine kinase receptor antibody
    see all

Images

  • Immunofluorescence analysis of TrkA expression in human SH-SY5Y (human neuroblastoma cell line from bone marrow) cells.
    Cells were fixed (4% formaldehyde, 10 minutes), permeabilized (0.1% Triton X-100) or non-permeabilized, and blocked (10% normal horse serum) for 30 minutes. SH-SY5Y cells were incubated for 1 hour with ab244209 at 2 μg/mL, (green). Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 μg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). Control cells were treated exactly the same way.

  • Immunofluorescence analysis of TrkA expression on SHSY-5Y (Human neuroblastoma cell line from bone marrow) cell membrane.

    Fixed (4% formaldehyde), non-permeabilized, and blocked (10% normal horse serum) SHSY-5Y cells were incubated with ab244209 at 2 μg/mL (green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-mouse-CF488A antibody (4 μg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue).

  • TrkA expression on non-permeabilized human SHSY-5Y (Human neuroblastoma cell line from bone marrow) and DU 145 (Human prostate carcinoma cell line) cells analysed by Flow Cytometry.
    Blocking: 200 μg/mL normal sheep IgG (30 minutes) on ice.
    Primary antibody: (ab244209, 2 μg per ~106 cells) for 30 minutes on ice.
    Secondary antibody: Goat anti-mouse PE (1/100 dilution, 20 min in dark on ice).
    Non-specific Control IgG clone X63 was used as negative control under same conditions (black dashed).

  • Comparison of TrkA expression on non-permeabilized and permeabilized human SHSY-5Y (Human neuroblastoma cell line from bone marrow) cells by Flow Cytometry.
    This batch of SHSY-5Y cells did not show TrkA expression on the membrane (left image), but immunoreactivity in permeabilized cells (right image), suggesting that TrkA is stored intracellularly.
    Conditions: Permeabilization with absolute methanol (10 mins on ice), or no permeabilization (incubation of primary antibody on ice only).
    Blocking: 200 μg/mL normal sheep IgG (30 mins) on ice.
    Primary antibody: (ab244209, 2 μg per ~106 cells) for 30 mins on ice.
    Secondary antibody: Goat anti-mouse PE (1:100 dilution, 20 min in dark on ice. Non-specific Control IgG, clone X63 was used as negative control under same conditions (black dashed).

References

ab244209 has not yet been referenced specifically in any publications.

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