Human NT-3 DuoSet ELISA

Catalog # Availability Size / Price Qty
DY267
Ancillary Products Available
Human NT-3 ELISA Standard Curve
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Product Details
Procedure
Citations (13)
FAQs
Supplemental Products
Reviews

Human NT-3 DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

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

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Neurotrophin-3. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

 

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
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

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: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

 

  

Data Example

Human NT-3 ELISA Standard Curve

Product Datasheets

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: NT-3

Neurotrophin-3 (NT-3) is a member of the NGF family of neurotrophic factors (also named neurotrophins) that are required for the differentiation and survival of specific neuronal subpopulations in both the central as well as the peripheral nervous systems. The neurotrophin family is comprised of at least four proteins including NGF, BDNF, NT-3, and NT-4/5. These secreted cytokines are synthesized as prepropeptides that are proteolytically processed to generate the mature proteins. All neurotrophins have six conserved cysteine residues that are involved in the formation of three disulfide bonds and all share approximately 55% sequence identity at the amino acid level. Similarly to NGF, bioactive NT-3 is predicted to be a noncovalently linked homodimer.

Human NT-3 cDNA encodes a 257 amino acid residue precursor protein with a signal peptide and a proprotein that are cleaved to yield the 119 amino acid residue mature NT-3. The amino acid sequence of mature NT-3 is identical in human, mouse, and rat. NT-3 transcripts have been detected in the cerebellum, hippocampus, placenta, heart, skin, and skeletal muscle. NT-3 primarily activates the TrkC tyrosine kinase receptor. In addition, NT-3 can activate Trk and TrkB kinase receptors in certain cell systems. NT-3 can also bind with low affinity to the low affinity NGF receptor.

Long Name:
Neurotrophin 3
Entrez Gene IDs:
4908 (Human)
Alternate Names:
HDNF; MGC129711; Nerve growth factor 2; Neurotrophic factor; neurotrophin 3; neurotrophin-3; NGF2; NGF-2; NT3; NT-3; NTF3

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

 

Citations for Human NT-3 DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

13 Citations: Showing 1 - 10
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  1. Descending motor circuitry required for NT-3 mediated locomotor recovery after spinal cord injury in mice
    Authors: Q Han, JD Ordaz, NK Liu, Z Richardson, W Wu, Y Xia, W Qu, Y Wang, H Dai, YP Zhang, CB Shields, GM Smith, XM Xu
    Nat Commun, 2019;10(1):5815.
    Species: Human
    Sample Types: Tissue Secretion
  2. NT-3/TrkC Axis Contributes to the Perineural Invasion and the Poor Prognosis in Human Salivary Adenoid Cystic Carcinoma
    Authors: H Li, Z Yang, W Wang, J Wang, J Zhang, J Liu, T Yang, Y Yang, J Wei, D Lei, X Yang
    J Cancer, 2019;10(24):6065-6073.
  3. Scaffold-Mediated Sustained, Non-viral Delivery of miR-219/miR-338 Promotes CNS Remyelination
    Authors: U Milbreta, J Lin, C Pinese, W Ong, JS Chin, H Shirahama, R Mi, A Williams, ME Bechler, J Wang, C Ffrench-Co, A Hoke, SY Chew
    Mol. Ther., 2018;0(0):.
  4. Transplantation of Recombinant Vascular Endothelial Growth Factor (VEGF)189-Neural Stem Cells Downregulates Transient Receptor Potential Vanilloid 1 (TRPV1) and Improves Motor Outcome in Spinal Cord Injury
    Authors: Y Zeng, H Han, B Tang, J Chen, D Mao, M Xiong
    Med. Sci. Monit., 2018;24(0):1089-1096.
    Species: Rat
    Sample Types: Cell Culture Supernates
  5. Serum levels of neurotrophic factors in active toxoplasmic retinochoroiditis
    Authors: Cynthia Azeredo Cordeiro
    Braz J Infect Dis, 2016;0(0):.
    Species: Human
    Sample Types: Serum
  6. Dynamic Response Genes in CD4+ T Cells Reveal a Network of Interactive Proteins that Classifies Disease Activity in Multiple Sclerosis
    Cell Rep, 2016;16(11):2928-39.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Improved axonal regeneration of transected spinal cord mediated by multichannel collagen conduits functionalized with neurotrophin-3 gene.
    Authors: Yao, L, Daly, W, Newland, B, Yao, S, Wang, W, Chen, B K K, Madigan, N, Windebank, A, Pandit, A
    Gene Ther, 2013;20(12):1149-57.
    Species: Human
    Sample Types: Tissue Homogenates
  8. Electrical stimulation of sympathetic neurons induces autocrine/paracrine effects of NGF mediated by TrkA.
    Authors: Saygili E, Schauerte P, Kuppers F, Heck L, Weis J, Weber C, Schwinger RH, Hoffmann R, Schroder JW, Marx N, Rana OR
    J. Mol. Cell. Cardiol., 2010;49(1):79-87.
    Species: Rat
    Sample Types: Cell Culture Supernates
  9. Improvement in disability after alemtuzumab treatment of multiple sclerosis is associated with neuroprotective autoimmunity.
    Authors: Jones JL, Anderson JM, Phuah CL, Fox EJ, Selmaj K, Margolin D, Lake SL, Palmer J, Thompson SJ, Wilkins A, Webber DJ, Compston DA, Coles AJ
    Brain, 2010;133(0):2232-47.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Insulin-like growth factor-1 and neurotrophin-3 gene therapy prevents motor decline in an X-linked adrenoleukodystrophy mouse model.
    Authors: Mastroeni R, Bensadoun JC, Charvin D, Aebischer P, Pujol A, Raoul C
    Ann. Neurol., 2009;66(1):117-22.
    Species: Human
    Sample Types: CSF
  11. Selected neurotrophins, neuropeptides, and cytokines: developmental trajectory and concentrations in neonatal blood of children with autism or Down syndrome.
    Authors: Nelson PG, Kuddo T, Song EY
    Int. J. Dev. Neurosci., 2006;24(1):73-80.
    Species: Human
    Sample Types: Complex Sample Type
  12. Neurotrophic factors in relapsing remitting and secondary progressive multiple sclerosis patients during interferon beta therapy.
    Authors: Caggiula M, Batocchi AP, Frisullo G, Angelucci F, Patanella AK, Sancricca C, Nociti V, Tonali PA, Mirabella M
    Clin. Immunol., 2005;118(1):77-82.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Controlled release of neurotrophin-3 from fibrin gels for spinal cord injury.
    Authors: Taylor SJ, McDonald JW, Sakiyama-Elbert SE
    J Control Release, 2004;98(2):281-94.
    Species: Human
    Sample Types: Protein

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