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.
Mouse IFN-gamma Quantikine ELISA Kit
R&D Systems | Catalog # MIF00
Loading...
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)
Loading...
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
Species
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.
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 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
Long Name
Interferon gamma
Alternate Names
IFG, IFI, IFNG, IFNgamma
Entrez Gene IDs
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 or batch 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
⚠ 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.Related Research Areas
Citations for Mouse IFN-gamma Quantikine ELISA Kit
Powered by Bioz
Customer Reviews for Mouse IFN-gamma Quantikine ELISA Kit (24)
4.8 out of 5
24 Customer Ratings
Have you used Mouse IFN-gamma Quantikine ELISA Kit?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Customer Images
Showing
1
-
5 of
24 reviews
Showing All
Filter By:
-
Sample Tested: Serum and PlasmaVerified Customer | Posted 02/25/2023
-
Sample Tested: EDTA PlasmaVerified Customer | Posted 11/15/2022
-
Sample Tested: Cell culture supernatantVerified Customer | Posted 10/26/2022Cell culture supernatants produced from transfected BHK-21 cells. samples regularly tested at much higher concentrations than the kit control, which was within range of acceptance, but was still very low.
-
Sample Tested: activated mouse CD8 T cellVerified Customer | Posted 09/18/2022Very good to use!
-
Sample Tested: mouse serumVerified Customer | Posted 06/26/2022Stable Kit! The R2 of the standard curve was more than 0.99.
-
Sample Tested: mouse serumVerified Customer | Posted 11/17/2021
-
Sample Tested: SerumVerified Customer | Posted 08/10/2021Standard curve was created based on assay protocol.
-
Sample Tested: SerumVerified Customer | Posted 10/14/2020
-
Sample Tested: activated mouse CD8 T cellVerified Customer | Posted 02/17/2020
-
Sample Tested: Cancer TissueVerified Customer | Posted 02/12/2020
-
Sample Tested: Caco-2 human colorectal adenocarcinoma cell lineVerified Customer | Posted 02/07/2020
-
Sample Tested: SerumVerified Customer | Posted 02/03/2020
-
Sample Tested: MDA-MD-231 human breast cancer cell lineVerified Customer | Posted 12/14/2019
-
Sample Tested: mouse pancreas tissue lysatesVerified Customer | Posted 10/30/2019
-
Sample Tested: Jurkat human acute T cell leukemia cell lineVerified Customer | Posted 04/04/2019
-
Sample Tested: activated mouse CD8 T cellVerified Customer | Posted 03/21/2019
-
Sample Tested: A172 human glioblastoma cell lineVerified Customer | Posted 05/30/2018
-
Sample Tested: SerumVerified Customer | Posted 05/30/2018
-
Sample Tested: SerumVerified Customer | Posted 03/23/2018We use this kit routinely in our lab with very consistent reliable results. Would certainly recommend!
-
Sample Tested: mouse serumVerified Customer | Posted 03/23/2018
-
Sample Tested: Cell culture supernatantVerified Customer | Posted 12/01/2017
-
Sample Tested: Concanavalin A-stimulated splenocytesVerified Customer | Posted 05/24/2017
-
Sample Tested: RAW 264.7 mouse monocyte/macrophage cell lineVerified Customer | Posted 05/02/2017
-
Sample Tested: Mouse primary CD8 cellsVerified Customer | Posted 04/27/2017Additional washes were added. Control or drug-treated CD8 cells were treated with allogeneic APC for 24 or 48 hours. Supernatants were subjected to IFN-g ELISA per manufacture's recommendation.
There are no reviews that match your criteria.
Protocols
View specific protocols for Mouse IFN-gamma Quantikine ELISA Kit (MIF00):
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.
- Add 50 µL of Assay Diluent to each well.
- Add 50 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 4 times for a total of 5 washes.
- Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
Loading...
Associated Pathways
Hematopoietic Stem Cell Differentiation Pathways & Lineage-specific Markers
IL-15 Signaling Pathways and their Primary Biological Effects in Different Immune Cell Types
IL-21 Signaling Pathways and their Primary Biological Effects in Different Immune Cell Types
Innate Lymphoid Cell Differentiation Pathways
