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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Mouse IFN-gamma DuoSet ELISA
R&D Systems | Catalog # DY485
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Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
31.2-2000 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
Mouse
Mouse 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 Mouse 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 mouse IFN-gamma. The suggested diluent is suitable for the analysis of most cell culture supernate 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)
Sample Volume Required
100 µL
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Scientific Data Images for Mouse IFN-gamma DuoSet ELISA
Mouse IFN-gamma ELISA Standard Curve
Kit Contents for Mouse 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 Mouse 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 Mouse IFN-gamma DuoSet ELISA
For research use only
Related Research Areas
Citations for Mouse IFN-gamma DuoSet ELISA
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Customer Reviews for Mouse IFN-gamma DuoSet ELISA (20)
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20 Customer Ratings
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Sample Tested: activated mouse CD8 T cellVerified Customer | Posted 01/24/2023
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Sample Tested: Mouse splenocytesVerified Customer | Posted 10/09/2020
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Sample Tested: Pancreatic cancer cellsVerified Customer | Posted 09/10/2020
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Sample Tested: Cell culture supernatantVerified Customer | Posted 06/11/2020We measured the INF-gamma level of culture supernatants from mouse spleen cells treated by group A or group B reagents. The data was normalized by comparing with that in control cells. The kit worked great and very sensitive! We loved it.
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Sample Tested: Breast cancer cellsVerified Customer | Posted 02/13/2020
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Sample Tested: BW5147 mouse T lymphoma cell lineVerified Customer | Posted 04/12/2018
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Sample Tested: activated mouse CD8 T cellVerified Customer | Posted 03/30/2018
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Sample Tested: Balb/cVerified Customer | Posted 03/28/2018
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Sample Tested: B16-F1 mouse melanoma cell lineVerified Customer | Posted 03/15/2018
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Sample Tested: 7TD1 mouse hybridoma cell lineVerified Customer | Posted 03/15/2018
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Sample Tested: MOUSE LUNGSVerified Customer | Posted 12/16/2017
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Sample Tested: LymphocytesVerified Customer | Posted 12/11/2017
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Sample Tested: Mouse heartVerified Customer | Posted 12/04/2017
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Sample Tested: MOUSE LUNGVerified Customer | Posted 11/27/2017
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Sample Tested: Brain tissueVerified Customer | Posted 07/21/2017
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Sample Tested: SerumVerified Customer | Posted 05/16/2017
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Sample Tested: Cell culture supernatantVerified Customer | Posted 09/01/2016
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Sample Tested: Serum and Plasma and Cell culture supernatantVerified Customer | Posted 03/23/2016You may use this kit with serum samples diluted >1:10.
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Sample Tested: Lymphocytes (ex vivo)Verified Customer | Posted 10/26/2015Specificity: Specific<br />Sensitivity: Sensitive<br />Buffer: Reagent Diluent 0.1% BSA,5 0.05% Tween 20 in Tris-buffered Saline (20 mM Trizma base, 150 mM NaCl), pH 7.2 - 7.4, filtered.<br />Dilution: 1/180
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Sample Tested: Lymphocytes (ex vivo)Verified Customer | Posted 10/26/2015Specificity: Specific<br />Sensitivity: Sensitive<br />Buffer: reagent diluent<br />Dilution: 100ng/ml (detection)
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Protocols
View specific protocols for Mouse IFN-gamma DuoSet ELISA (DY485):
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
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- 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
