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Human Total TrkA DuoSet IC ELISA, 2 Plate 1 KT

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Human Total TrkA DuoSet IC ELISA, 2 Plate 1 KT信息二維碼

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產(chǎn)品介紹

    基本參數(shù)

    詳細(xì)說明

    • Assay Type

      Solid Phase Sandwich ELISA

    • Format

      96-well strip plate

    • Assay Length

      4 hours 40 mins (after plate preparation)

    • Sample Type & Volume Required

      Cell lysates (100 μL)

    • Range

      156.00 - 10,000 pg/mL

    • Sufficient Materials

      Kits available for two, five, or fifteen 96-well plates*

    • Specificity

      Please see the

    * Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.



     

    Product Features

    • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time

    • Development protocols are provided to guide further assay optimization

    • Assay can be customized to your specific needs

    • Available in 2, 5, and 15- (96-well) plate pack sizes

    • Economical alternative to Western blot

    Kit Content

    • Capture Antibody

    • Conjugated Detection Antibody

    • Calibrated Immunoassay Standard or Control

    • Streptavidin-HRP

    Other Reagents Required


      PBS: (Catalog # ), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na  2HPO  4, 1.5 mM KH  2O  4, pH 7.2 - 7.4, 0.2 μm filtered  
     
      Wash Buffer: (Catalog # ), or equivalent  
     
      Lysis Buffer*  
     
      IC Diluent*

    Blocking Buffer*
     
     
      Substrate Solution: 1:1 mixture of Color Reagent A (H  2O  2) and Color Reagent B (Tetramethylbenzidine) (Catalog # )  
     
      Stop Solution: 2 N H  2SO  4 (Catalog # )  
     
      Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # ), or equivalent  
     
      Plate Sealers: ELISA Plate Sealers (Catalog # ), or equivalent

    *For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

    Data Examples


    Figure 1: The Human Total TrkA DuoSet? IC ELISA is more sensitive than immunoprecipitation (IP)-Western blot analysis. Lysates prepared from C6 rat glioma cells transfected with human TrkA (C6-hTrkA) were serially diluted and analyzed by (A) IP-Western blot and (B) this DuoSet? IC ELISA. IPs were performed using an anti-TrkA monoclonal antibody and goat anti-mouse agarose. Immunoblots were incubated with a biotinylated anti-TrkA polyclonal antibody to detect total TrkA. Bands were visualized with Streptavidin-HRP (Catalog # ) followed by chemiluminescent detection. Human TrkA can be detected by this Human Total TrkA DuoSet? IC ELISA by using approximately 2 to 4 times less lysate than is needed for a conventional IP-Western Blot.

    Preparation and Storage

    • Storage

      Store the unopened product at 2 - 8 °C. Do not use past expiration date.

    Background: TrkA

    TrkA, the product of the protooncogene trk, is a member of the neurotrophic tyrosine kinase receptor family that has three members. TrkA, TrkB and TrkC preferentially bind NGF, NT-4 and BDNF, and NT-3, respectively. 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.

    Two distinct TrkA isoforms that differ by virtue of a 6 amino acid insertion in their extracellular domain have been identified. The longer TrkA isoform is the only isoform expressed within neuronal tissues whereas the shorter TrkA is expressed mainly in non neuronal tissues. NGF binds to TrkA with low affinity and activates its cytoplasmic kinase, initiating a signaling cascade that mediates neuronal survival and differentiation. Higher affinity binding of NGF requires the coexpression of TrkA with the p75 NGF receptor (NGFR), a member of the tumor necrosis factor receptor superfamily. NGFR binds all neurotrophins with low affinity and modulates Trk activity as well as alters the specificity of Trk receptors for their ligands. NGFR can also mediate cell death when expressed independent of Trk.

    • Entrez Gene IDs:

      4914 (Human); 59109 (Rat);

    • Long Name:

      Neurotrophic Tyrosine Kinase Receptor A

    • Aliases:

      DKFZp781I14186; EC 2.7.10; EC 2.7.10.1; MTChigh affinity nerve growth factor receptor; Neurotrophic tyrosine kinase receptor type 1; neurotrophic tyrosine kinase, receptor, type 1; NTRK1; NTRK-1; p140-TrkA; TRK1-transforming tyrosine kinase protein; TrkA; Trk-A; TRKAOncogene TRK; TRKTRK1; tyrosine kinase receptor A

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