Skip to main content

Human Total TrkA DuoSet IC ELISA

R&D Systems, part of Bio-Techne | Catalog # DYC175-2

R&D Systems, part of Bio-Techne
Discontinued Product
DYC175-2 has been discontinued. View all TrkA products.

Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

156-10000 pg/mL

Sample Type

Cell Lysates

Reactivity

Human

Human Total TrkA DuoSet IC ELISA 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

Product Summary for Human Total TrkA DuoSet IC ELISA

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total TrkA in cell lysates. An immobilized capture antibody specific for TrkA binds both phosphorylated and unphosphorylated TrkA. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

Cell lysates (100 µL)

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Label

HRP

Scientific Data Images for Human Total TrkA DuoSet IC ELISA

Standard Curve

The Human Total TrkA DuoSet® IC ELISA is more sensitive than immunoprecipitation (IP)-Western blot analysis.

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 # DY998) 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.

Kit Contents for Human Total TrkA DuoSet IC ELISA

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control
  • Streptavidin-HRP

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*


Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

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

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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.

Long Name

Neurotrophic Tyrosine Kinase Receptor A

Alternate Names

NTRK-1, NTRK1

Entrez Gene IDs

4914 (Human); 18211 (Mouse); 59109 (Rat); 102129364 (Cynomolgus Monkey)

Gene Symbol

NTRK1

Additional TrkA Products

Product Documents for Human Total TrkA DuoSet IC ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Total TrkA DuoSet IC ELISA

For research use only

Loading...
Loading...
Loading...