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.
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Human IFN-gamma DuoSet ELISA
R&D Systems | Catalog # DY285B
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Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
9.38-600 pg/mL
Sample Type
Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet
Reactivity
Human
Human IFN-gamma DuoSet ELISA Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
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Product Summary for Human IFN-gamma DuoSet ELISA
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Interferon gamma (IFN-γ). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Specifications
Assay Format
96-well strip plate (sold separately)
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Human IFN-gamma DuoSet ELISA
Human IFN-gamma ELISA Standard Curve
Kit Contents for Human IFN-gamma DuoSet ELISA
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008C) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Reagent Diluent*
Blocking Buffer*
Substrate Solution: ELISA TMB Substrate (Catalog # DY999B or DY999B-250)
Stop Solution: Methanesulfonic acid (Catalog # DY994B or DY994B-250)
Microplates: (Catalog # DY990), or equivalent
Plate Sealers: (Catalog # DY992), or equivalent
*For the recommended Reagent Diluent and Blocking Buffer for a specific DuoSet ELISA Development Kit, refer to the product datasheet.
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 Human IFN-gamma DuoSet ELISA
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 IFN-gamma DuoSet ELISA
For research use only
Related Research Areas
Citations for Human IFN-gamma DuoSet ELISA
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Customer Reviews for Human IFN-gamma DuoSet ELISA (15)
4.4 out of 5
15 Customer Ratings
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Sample Tested: EDTA PlasmaVerified Customer | Posted 02/28/2025EDTA plasma, used neat
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Sample Tested: Cell culture supernatant and THP-1 human acute monocytic leukemia cell lineVerified Customer | Posted 09/05/2024
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Sample Tested: PlasmaVerified Customer | Posted 11/16/2022Plasma tested at 2-fold dilution but appears to cause a matrix effect.
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Sample Tested: Natural killer cellsVerified Customer | Posted 10/10/2022
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Sample Tested: Cell culture supernatantVerified Customer | Posted 10/13/2021
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Sample Tested: Cell culture supernatantVerified Customer | Posted 09/28/2021
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Sample Tested: Tissue Culture MediaVerified Customer | Posted 09/29/2020Detection of IFN-g produced by BCMA-specific CAR-T cells in response to BCMA-negative and BCMA-positive hematopoietic cell lines.
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Sample Tested: T cellsVerified Customer | Posted 08/26/2020
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Sample Tested: Blood mononuclear cells (PBMCs)Verified Customer | Posted 06/16/2020
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Sample Tested: cell supernatantVerified Customer | Posted 12/12/2019
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Sample Tested: EDTA PlasmaVerified Customer | Posted 07/02/2019
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Sample Tested: MOLT-4 human acute lymphoblastic leukemia cell line and Jurkat human acute T cell leukemia cell lineVerified Customer | Posted 05/01/2019
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Sample Tested: Cell culture supernatantVerified Customer | Posted 12/17/2018
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Sample Tested: Natural killer cells and Cell culture supernatantVerified Customer | Posted 11/16/2018Nice linear curve. Good detection rate for the tissue culture supernatant
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Sample Tested: SerumVerified Customer | Posted 11/21/2017We used human serum at a dilution of 1:1 with an good technical result. However, IFNg was not detectable in human serum in our conditions.
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Protocols
View specific protocols for Human IFN-gamma DuoSet ELISA (DY285B):
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL of Block Buffer to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
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
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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
