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Mouse IFN-gamma ELISA Kit - Quantikine

R&D Systems, part of Bio-Techne | Catalog # MIF00

R&D Systems, part of Bio-Techne
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MIF00-1
PMIF00
SMIF00

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL)

Sensitivity

2 pg/mL

Assay Range

9.4-600 pg/mL (Cell Culture Supernates, Serum)

Product Summary for Mouse IFN-gamma Quantikine ELISA Kit

The Quantikine Mouse IFN-gamma Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IFN-gamma in cell culture supernates and mouse serum. It contains E. coli-expressed mouse IFN-gamma and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant mouse IFN-gamma accurately. Results obtained using natural mouse IFN-gamma showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IFN-gamma.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Mouse

Specificity

Natural and recombinant mouse 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.

Sample Values

Serum - Forty individual mouse serum samples were evaluated for the presence of mouse IFN-gamma in this assay. Thirty-eight samples measured less than the lowest Mouse IFN-g Standard, 9.4 pg/mL. Two samples read 24 pg/mL and 60 pg/mL, respectively.

Cell Culture Supernates: Mouse splenocytes (2 x 106 cells/mL) were cultured for 3 days in RPMI plus 10% fetal bovine serum and stimulated with 10 μg/mL PHA. An aliquot of the cell culture supernate was removed, assayed for mouse IFN-gamma, and measured 32 ng/mL. 

EL-4 mouse lymphoblast cells (9 x 105 cells/mL) were cultured for 2 days in DMEM plus 10% fetal bovine serum and stimulated with 10 μg/mL PHA and 10 ng/mL PMA. An aliquot of the cell culture supernate was removed, assayed for mouse IFN-gamma, and measured 4 ng/mL.

Mouse lung conditioned media (1 lung, 1-2 mm pieces in 10 mL of medium) was collected after culturing for 5 days in RPMI plus 10% fetal bovine serum. An aliquot of the cell culture supernate was removed, assayed for mouse IFN-gamma, and measured 198 pg/mL.

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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 44.8 156 518 43.3 144 509
Standard Deviation 2.2 5.0 14.2 3.6 14.7 48.6
CV% 4.9 3.2 2.7 8.3 10.2 9.5

Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 45.1 143 528 42.8 139 518
Standard Deviation 2.1 4.4 11.8 3.6 13.6 46.5
CV% 4.7 3.1 2.2 8.4 9.8 9.0

Recovery for Mouse IFN-gamma Quantikine ELISA Kit

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=7) 105 98-115
Serum (n=5) 97 91-106

Linearity

To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse IFN-gamma in each matrix were diluted with the appropriate Calibrator Diluent and then assayed.

Mouse IFN-gamma ELISA Linearity

Scientific Data Images for Mouse IFN-gamma Quantikine ELISA Kit

Mouse IFN-gamma ELISA Calibrator Diluent RD5Y Standard Curve

Mouse IFN-gamma ELISA Calibrator Diluent RD5Y Standard Curve

Mouse IFN-gamma ELISA Calibrator Diluent RD6-12 Standard Curve

Mouse IFN-gamma ELISA Calibrator Diluent RD6-12 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: 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 Mouse IFN-gamma Quantikine ELISA Kit

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse IFN-gamma Quantikine ELISA Kit

For research use only

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⚠ 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.