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.
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
The recovery of CNTF spiked to three different levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=5)
Citrate Plasma (n=5)
EDTA Plasma (n=5)
Heparin Plasma (n=5)
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.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
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.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
50 µL Assay Diluent
Add 50 µL of Assay Diluent to each well.
200 µL Standard, Control, or Sample
Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
Aspirate each well and wash, repeating the process twice for a total of 3 washes.
200 µL Conjugate
Add 200 µL of Conjugate to each well.
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.
Aspirate and wash 3 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
50 µL Stop Solution
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.
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