• Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours or 4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (200 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL), Citrate Plasma (200 uL)
  • Sensitivity
    8 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
  • Specificity
    Recombinant human CNTF
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC20, Quantikine Immunoassay Control Group 3 - Please Inquire
Product Summary
The Quantikine Human CNTF Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure recombinant human CNTF in cell culture supernates, serum, and plasma. It contains recombinant human CNTF and has been shown to accurately quantitate the recombinant factor.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean1104611295128399821
Standard Deviation510.231.410.126.852.6
CV%4.52.22.47.96.76.4

Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean10340612012066711388
Standard Deviation4.41228.219.647.893.9
CV%4.332.39.57.16.8

Recovery

The recovery of CNTF spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 99 92-115
Citrate Plasma (n=5) 87 71-125
EDTA Plasma (n=5) 90 78-97
Heparin Plasma (n=5) 92 82-98
Serum (n=5) 95 85-101
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of CNTF in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human CNTF Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: CNTF
Ciliary neurotrophic factor (CNTF) is structurally related to IL-6, IL-11, LIF, CLC, and OSM. CNTF was initially identified as a trophic factor for embryonic chick ciliary parasympathetic neurons in culture. Subsequent studies have demonstrated that CNTF is a survival factor for additional neuronal cell types, including dorsal root ganglion sensory neurons, sympathetic ganglion neurons, embryonic motor neurons, major pelvic ganglion neurons and hippocampal neurons.
    • Long Name
      Ciliary Neurotrophic Factor
    • Entrez Gene IDs
      1270 (Human); 25707 (Rat);
    • Alternate Names
      ciliary neurotrophic factor; CNTF; HCNTF;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 50 µL Assay Diluent
    4.   Add 50 µL of Assay Diluent to each well.

    5. 200 µL Standard, Control, or Sample
    6.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    7.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well.
    10.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
      For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    11.   Aspirate and wash 3 times.

    12. 200 µL Substrate Solution
    13.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.

    14. 50 µL Stop Solution
    15. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    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.

    7 Citations: Showing 1 - 7
    Filter your results:

    Species
    Sample Type
    1. Purification and characterization of a long-acting ciliary neurotrophic factor via genetically fused with an albumin-binding domain
      Authors: L Xu, C Zhang, L Liu, Y Zhang, Q Wang, J Wang, Y Liu, Z Su
      Protein Expr. Purif., 2017;0(0):.
      Species: Rat
      Sample Type: Serum
    2. Ciliary neurotrophic factor (CNTF) for human retinal degeneration: phase I trial of CNTF delivered by encapsulated cell intraocular implants.
      Authors: Sieving PA, Caruso RC, Tao W, Coleman HR, Thompson DJ, Fullmer KR, Bush RA
      Proc. Natl. Acad. Sci. U.S.A., 2006;103(10):3896-901.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. Transplantation of transduced retinal pigment epithelium in rats.
      Authors: Saigo Y, Abe T, Hojo M, Tomita H, Sugano E, Tamai M
      Invest. Ophthalmol. Vis. Sci., 2004;45(6):1996-2004.
      Species: Rat
      Sample Type: Cell Culture Supernates
    4. AF5, a CNS cell line immortalized with an N-terminal fragment of SV40 large T: growth, differentiation, genetic stability, and gene expression.
      Authors: Truckenmiller ME, Vawter MP, Zhang P, Conejero-Goldberg C, Dillon-Carter O, Morales N, Cheadle C, Becker KG, Freed WJ
      Exp. Neurol., 2002;175(0):318.
      Species: Rat
      Sample Type: Cell Culture Supernates
    5. Restoration of cognitive and motor functions by ciliary neurotrophic factor in a primate model of Huntington's disease.
      Authors: Mittoux V, Joseph JM, Conde F, Palfi S, Dautry C, Poyot T, Bloch J, Deglon N, Ouary S, Nimchinsky EA, Brouillet E, Hof PR, Peschanski M, Aebischer P, Hantraye P
      Hum. Gene Ther., 2000;11(8):1177-87.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Cytokine production in rhesus monkeys infected with Plasmodium coatneyi.
      Authors: Yang C, Xiao L, Tongren JE, Sullivan J, Lal AA, Collins WE
      Am. J. Trop. Med. Hyg., 1999;61(2):226-9.
      Species: Primate
      Sample Type: Serum
    7. Regional changes of ciliary neurotrophic factor and nerve growth factor levels in post mortem spinal cord and cerebral cortex from patients with motor disease.
      Authors: Anand P, Parrett A, Martin J, Zeman S, Foley P, Swash M, Leigh PN, Cedarbaum JM, Lindsay RM, Williams-Chestnut RE
      Nat. Med., 1995;1(2):168-72.
      Species: Human
      Sample Type: Tissue Homogenates
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Group 3

    Ctrl QC20
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    Supplemental ELISA Products
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    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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