製品の概要

  • 製品名Anti-TrkA antibody [EP1058Y]
    TrkA 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP1058Y] to TrkA
  • 特異性This antibody detects both phosphorylated and unphosphorylated TrkA.
  • アプリケーション適用あり: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human TrkA. A synthetic peptide corresponding to residues surrounding tyrosine 791 of human TrkA.

  • ポジティブ・コントロール
    • WB: Human, mouse and rat brain tissue lysates. IHC-P: Human liver and brain tissue. ICC/IF: U87-MG and SH-SY5Y cells. Flow Cyt: SH-SY5Y cells. IP: Rat brain tissue lysate.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

     

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

     

    Available versions:

    Anti-TrkA antibody (Alexa Fluor® 488) [EP1058Y] (ab194321)

    Anti-TrkA antibody (Alexa Fluor® 647) [EP1058Y] (ab194322)

    Anti-TrkA antibody (HRP) [EP1058Y] (ab194323)

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab76291 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ICC/IF 1/100.
WB 1/1000 - 1/10000. Predicted molecular weight: 87 kDa.
IP 1/40.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt 1/130.

(paraformaldehyde or methanol fixed cells)

Permeabilization recommended



ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能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.
  • 組織特異性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.
  • 関連疾患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).
  • 配列類似性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.
  • ドメインThe transmembrane domain mediates interaction with KIDINS220.
    The extracellular domain mediates interaction with NGFR.
  • 翻訳後修飾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.
  • 細胞内局在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
  • 参照データベース
  • 別名
    • 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

Anti-TrkA antibody [EP1058Y] 画像

  • Anti-TrkA antibody [EP1058Y] (ab76291) at 1/1000 dilution (purified) + Human brain tissue lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 87 kDa
    Observed band size : 145 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-TrkA antibody [EP1058Y] (ab76291) at 1/5000 dilution (purified) + Rat brain tissue lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 87 kDa
    Observed band size : 145 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-TrkA antibody [EP1058Y] (ab76291) at 1/5000 dilution (purified) + Mouse brain tissue lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 87 kDa
    Observed band size : 145 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-TrkA antibody [EP1058Y] (ab76291) at 1/5000 dilution (unpurified)

    Lane 1 : Rat brain tissue lysate (untreated)
    Lane 2 : Rat brain tissue lysate (treated with AP)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
    Lane 2 : HRP-conjugate goat anti-rabbit IgG at 1/2000 dilution


    Predicted band size : 87 kDa
    Observed band size : 145 kDa (why is the actual band size different from the predicted?)
  • All lanes : Anti-TrkA antibody [EP1058Y] (ab76291) at 1/2500 dilution (unpurified)

    Lane 1 : Human neuronal cell lysate (untreated)
    Lane 2 : Human neuronal cell lysate (treated with rh-ßNGF at 25ng/ml)
    Lane 3 : Human neuronal cell lysate (treated with rh-ßNGF at 50ng/ml)
    Lane 4 : Human neuronal cell lysate (treated with rh-NT3 at 50ng/ml)
    Lane 5 : Human neuronal cell lysate (treated with rh-BDNF 50ng/ml)
    Lane 6 : TrkA protein

    Lysates/proteins at 15 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit polyclonal IgG at 1/2000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 87 kDa
    Observed band size : 145 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling TrkA with purified ab76291 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue labelling TrkA with unpurified ab76291 at 1/100 dilution.

  • Unpurified ab76291 staining TrkA in murine brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed with formaldehyde, permeabilized with 0.1% Saponin/PBS and blocked with 4% serum for 30 minutes at 25°C, antigen retrieval was by heat mediation with a citrate buffer. Samples were incubated with primary antibody (1/150 in blocking buffer) for 16 hours at 4°C. A FITC-conjugated goat anti-rabbit polyclonal IgG (1/100) was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/Immunofluorescence analysis of U87-MG cells labelling TrkA with purified ab76291 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/100) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).

  • Immunocytochemsitry/Immunofluorescence analysis of SH-SY5Y cells labelling TrkA with unpurified ab76291 at a 1/100 dilution.

  • ICC/IF image of TrkA staining on culture of mouse DRG neurons using unpurified ab76291 (1/100). The cells were fixed using formaldehyde and permeabilized using 0.2% Triton X-100. The cells were blocked using 10% Goat serum for 1 hour at 22°C. Unpurified ab76291 was diluted 1/100 using PBS and incubated with the cells for 30 mins at 22°C. The secondary antibody used was Goat polyclonal to Rabbit IgG conjugated to Alexa Fluor® 488 (1/1000). Neuron was stained using Beta III tubulin antibody (Alexa Fluor® 647)

    See Abreview

  • Overlay histogram showing SH-SY5Y cells stained with unpurified ab76291 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76291, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with methanol (5 min) used under the same conditions.
    Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

  • Flow cytometry analysis of SH-SY5Y cells labelling TrkA with purified ab76291 at 1/130 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

  • ab76291 (purified) at 1/40 immunoprecipitating TrkA in rat brain tissue lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

Anti-TrkA antibody [EP1058Y] (ab76291) 使用論文

This product has been referenced in:
  • Tatematsu T  et al. Investigation of neurotrophic tyrosine kinase receptor 1 fusions and neurotrophic tyrosine kinase receptor family expression in non-small-cell lung cancer and sensitivity to AZD7451 in vitro. Mol Clin Oncol 2:725-730 (2014). IHC-P ; Human . Read more (PubMed: 25054037) »
  • Zhao T  et al. Levo-tetrahydropalmatine retards the growth of ectopic endometrial implants and alleviates generalized hyperalgesia in experimentally induced endometriosis in rats. Reprod Sci 18:28-45 (2011). Rat . Read more (PubMed: 20884991) »

See all 2 Publications for this product

Product Wall

Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Nasal mucosa)
Specification Nasal mucosa
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative Paraformaldehyde
Username

Mr. Duc Dung Le

Verified customer

投稿 Jun 09 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (breast cancer, fetal liver)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA, pH9, 100C, 30 min
Permeabilization No
Specification breast cancer, fetal liver
Blocking step H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Jun 11 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer ph=6
Sample Mouse Tissue sections (Epithelium in the nasal fossae)
Specification Epithelium in the nasal fossae
Permeabilization No
Fixative Formaldehyde
Username

Dr. Francois Daubeuf

Verified customer

投稿 Mar 11 2015

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (Precast Gel 4-12%, Criterion TGX (Biorad))
Sample Mouse Tissue lysate - whole (Brain)
Specification Brain
Treatment Tissue was mixed in Rippa buffer and denaturated in Laemmli buffer 5min ß 95¯C
Blocking step Milk as blocking agent for 1 minute(s) · Concentration: 0.5% · Temperature: 4°C
Username

Dr. Francois Daubeuf

Verified customer

投稿 Mar 05 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Mouse Cell (DRG neuron)
Specification DRG neuron
Fixative Formaldehyde
Permeabilization Yes - 0.2% Triton X-100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Username

Dr. Franziska Denk

Verified customer

投稿 May 02 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (neuronal cells)
Loading amount 15 µg
Specification neuronal cells
Treatment see below (Additional notes)
Gel Running Conditions Reduced Non-Denaturing (Native) (5-16% gels)
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Aug 09 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Rat Cell (PC12 cells (neuronal cells))
Specification PC12 cells (neuronal cells)
Preparation Cell harvesting/tissue preparation method: cell culture (commercially available cell line)
Sample buffer: 0,1% Saponin /1% FCS/ PBS
Fixation Paraformaldehyde
Permeabilization Yes - 0,1% Saponin / PBS
Gating Strategy no gate
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Abcam user community

Verified customer

投稿 Aug 09 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (mouse brain)
Specification mouse brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer, pH 6.0
Permeabilization Yes - 0,1% Saponin / PBS
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Aug 09 2012

Thank you for your inquiry.

We recommend to incubate primary antibodies at 4C over night for a standard IHC protocol. This will ensure the most specific binding and the best saturation with the antibody.

For more information on IHC ...

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Thank you for your inquiry.

ab1840:

Which serum is used as blocking agent is dependent on the secondary antibody that is used. The serum should be from the same species in which the secondary antibody was produced.

For exa...

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