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Human CNTF Quantikine ELISA Kit

R&D Systems | Catalog # DNT00

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Key Product Details

Assay Length

3.5 hours or 4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (200 µL), Serum (200 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL), Citrate Plasma (200 µL)

Sensitivity

8 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human CNTF Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

精度

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
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 110 461 1295 128 399 821
Standard Deviation 5.0 10.2 31.4 10.1 26.8 52.6
CV% 4.5 2.2 2.4 7.9 6.7 6.4

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 103 406 1201 206 671 1388
Standard Deviation 4.4 12.0 28.2 19.6 47.8 93.9
CV% 4.3 3.0 2.3 9.5 7.1 6.8

Recovery for Human CNTF Quantikine ELISA Kit

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 ELISA Linearity

Scientific Data Images for Human CNTF Quantikine ELISA Kit

Human CNTF ELISA Cell Culture Supernate Standard Curve

Human CNTF ELISA Cell Culture Supernate Standard Curve

Human CNTF ELISA Serum/Plasma Standard Curve

Human CNTF ELISA Serum/Plasma Standard Curve

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: 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

Alternate Names

ciliary neurotrophic factor, HCNTF

Entrez Gene IDs

1270 (Human); 12803 (Mouse); 25707 (Rat)

Gene Symbol

CNTF

Additional CNTF Products

Product Documents for Human CNTF Quantikine ELISA Kit

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human CNTF Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human CNTF Quantikine ELISA Kit

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    Name: Anonymous
    Sample Tested: Human cell conditioned medium
    Verified Customer | Posted 12/03/2018

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Protocols

View specific protocols for Human CNTF Quantikine ELISA Kit (DNT00):

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
  3.   Add 50 µL of Assay Diluent to each well.
    4. 200 µL Standard, Control, or Sample
  4.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well.
  7.   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.
  8.   Aspirate and wash 3 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    10. 50 µL Stop Solution
  10. 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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