Canine IFN-gamma Quantikine ELISA Kit

R&D Systems | Catalog # CAIF00

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Key Product Details

Assay Length

4 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)

Sensitivity

60 pg/mL

Assay Range

62.5-4000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Canine IFN-gamma Quantikine ELISA Kit

The Quantikine Canine IFN-gamma Immunoassay is a 4 hour solid-phase ELISA designed to measure canine IFN-gamma in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant canine IFN-gamma and antibodies raised against the recombinant factor. Results obtained for naturally occurring canine IFN-gamma showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural canine IFN-gamma.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

Biotin

Species

Canine

Specificity

Natural and recombinant canine IFN-gamma

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.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times 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, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 111 459 1234 119 457 1315
Standard Deviation 10.1 35.1 92.4 10.1 21.5 64.8
CV% 9.1 7.6 7.5 8.5 4.7 4.9

Recovery for Canine IFN-gamma Quantikine ELISA Kit

The recovery of canine IFN-gamma spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 103 92-111
EDTA Plasma (n=4) 96 86-102
Heparin Plasma (n=4) 98 93-106
Serum (n=4) 96 83-107

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of canine IFN-gamma in each matrix were diluted with Calibrator Diluent and assayed.

Canine IFN-gamma ELISA Linearity

Scientific Data Images for Canine IFN-gamma Quantikine ELISA Kit

Canine IFN-gamma ELISA Standard Curve

Detection of Canine IFN-gamma by ELISA

Detection of Canine IFN-gamma by ELISA

Serum IFN-gamma induction was associated with NHS-IL12 dose and the observation of adverse events.IFN-gamma levels were measured using ELISA techniques. Induction of IFN-gamma was detectable in dogs treated with NHS-IL12 at a dose of 1.6 mg/m2 or higher. IFN-gamma levels spiked sharply at 24 hours post-treatment and returned to undetectable levels by 48 hours. Elevated serum IFN-gamma (> 100 pg/ml) was associated with increased risk for toxicity. However the highest level of IFN-gamma induction was not directly linked to the most severe (Grade 4 or 5) adverse events. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26091536), licensed under a CC-BY license. Not internally tested by R&D Systems.

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: IFN-gamma

IFN-gamma (Interferon-gamma) is the prototype proinflammatory cytokine and is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells. It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, upregulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects. In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation. IFN-gamma dimers signal through a receptor complex of two IFN-gamma R1 and two IFN-gamma R2 subunits.

Long Name

Interferon gamma

Alternate Names

IFG, IFI, IFNG, IFNgamma

Entrez Gene IDs

3458 (Human); 15978 (Mouse); 25712 (Rat); 396991 (Porcine); 281237 (Bovine); 403801 (Canine); 493965 (Feline)

Gene Symbol

IFNG

Additional IFN-gamma Products

Product Documents for Canine IFN-gamma 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 Canine IFN-gamma 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 Canine IFN-gamma Quantikine ELISA Kit

Customer Reviews for Canine IFN-gamma Quantikine ELISA Kit (5)

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  • Canine IFN-gamma Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: cell supernatant from cultured T cells
    Verified Customer | Posted 06/16/2020
    Canine IFN-gamma Quantikine ELISA Kit CAIF00
  • Canine IFN-gamma Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell Culture Media
    Verified Customer | Posted 05/26/2020
    The standards in this kit work very well - over 0.99 R squared every time I use it. Great assay. Would recommend to a colleague. Consistent and reliable.
    Canine IFN-gamma Quantikine ELISA Kit CAIF00
  • Canine IFN-gamma Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Peripheral blood mononuclear cells (PBMCs)
    Verified Customer | Posted 03/10/2020
  • Canine IFN-gamma Quantikine ELISA Kit
    Name: Leslie Priddy
    Sample Tested: Serum
    Verified Customer | Posted 04/03/2018
  • Canine IFN-gamma Quantikine ELISA Kit
    Name: chen xie
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 12/01/2017

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Protocols

View specific protocols for Canine IFN-gamma Quantikine ELISA Kit (CAIF00):

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. 50 µL Standard, Control, or sample
  4.   Add 50 µL of Standard, Control, or sample to each well. Tap the plate gently for 1 minute. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    6. 100 µL Conjugate
  6.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  7.   Aspirate and wash 5 times.
    8. 100 µL Streptavidin-HRP
  8.   Add 100 µL Streptavidin-HRP to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
  9.   Aspirate and wash 5 times.
    10. 100 µL Substrate Solution
  10. Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    11. 100 µL Stop Solution
  11. Add 100 µL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Canine IFN-gamma Quantikine ELISA Kit

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  • Q: Assay Diluent RD1-63 contains visible precipitate. Is this acceptable to use?

    A: A small amount of precipitate is expected and acceptable in Assay Diluent RD1-63. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.

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