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Product Details
Product Overview
Human IFN-γ ELISA Kit, PRO is a ready-to-use immunoassay kit, specifically designed to quantitate natural and recombinant human IFN-gamma that is present in biological samples, such as human serum, plasma, and cell culture supernatants. Our ELISA Kit provides several benefits:
- Standards to calibrate with NIBSC/WHO standards for comparable results.
- Fully validation in biologic samples for detection range, sensitivity, inter- and intra-plate CV, recovery, dilution linearity, specificity, and matrix effects to ensure reliable results according to ICH M10 guideline.
- High-quality antibody pairs and protein standards, along with rigorous quality control, to guarantee consistent results across different batches.
- Simplified and straightforward protocols and ready-to-use reagents to save assay time.
Product Specifications
Assay TypeSandwich-ELISAAnalyteIFN-γFormat96-wells plate breakable into 12 x 8 wells stripsReactivityHumanSensitivity4 pg/mLRange7.81 pg/mL-1000 pg/mLAssay Time2 hrSample volume50 μLSample TypeCell Culture Supernatants, Plasma, Serum.NIBSC Code82/587Elevate your research experience with our Cytokine/Biomarker Detection Kits, where accuracy, reliability, and ease of use are converging to deliver exceptional results.
Application
The kit is developed for quantitative detection of natural and recombinant human IFN-gamma in serum, plasma and cell culture supernatants.
It is suitable for potency assay of CAR-T/NK cell therapy. It has been calibrated against a highly purified human IFN-γ and is evaluated with standard from NIBSC/WHO. Reference Reagent INTERFERON GAMMA (Human, rDNA derived) NIBSC code: 82/587.
It is for research use only.
Workflow
Storage
Keep the unopened kit stored at 2-8 °C. Avoid using the kit beyond its expiration date. The opened kit should be stored per components table. The shelf life is 30 days from the date of opening.
Materials Provided
IDComponentsSizeCEA006-C01Pre-coated Anti-IFN-γ Antibody Microplate1 plateCEA006-C02Human IFN-γ Standard14 μg/48 tests
14 μg×2/96 testsCEA006-C03Biotin-Anti-IFN-γ Antibody Con. Solution75 μL/48 tests
150 μL/96 testsCEA006-C04Biotin-Antibody Dilution Buffer8 mLCEA006-C05Streptavidin-HRP Con. Solution250 μL/48 tests
500 μL/96 testsCEA006-C06Streptavidin-HRP Dilution Buffer15 mLCEA006-C0720×Washing Buffer50 mLCEA006-C08Sample Dilution Buffer15 mL/48 tests
15 mL×2/96 testsCEA006-C09Substrate Solution12 mLCEA006-C10Stop Solution6 mLACRO Quality Management System
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Performance Data
Typical Data
Please refer to DS document for the assay protocol.
For each experiment, a standard curve needs to be set for each microplate, and the specific OD value may vary depending on different laboratories, testers, or equipment. The following example data is for reference only. The sample concentration was calculated based on the results of the standard curve. The minimum detectable concentration of IFN-γ is less than 4.0 pg/mL.
Sample Values
Serum Sample: IFN-γ concentrations were measured in two hunderd and eight human serum samples, which were collected from healthy human subjects. The measurements of two hunderd and eight samples are shown in the figure below.
Matrix Effect
Three types of materials were tested to observe if there were matrix effect (interference). If the concentration of hemoglobin (simulated hemolysis) is less than 3500 mg/dL, the concentration of triglyceride (simulated lipid blood) is less than 3.0 mg/mL, and the concentration of bilirubin (simulated jaundice) is less than 20 mg/dL, testing results will not be affected.
Hook Effect
According to the hook effect evaluation, tested results will not be affected when IFN-γ concentration is higher than 250 ng/mL.
Dilution Linearity
High concentrations of human IFN-γ serum samples were diluted with 1:2, 1:4, 1:8, 1:16, 1:32 and 1:64 ratios for gradient dilution to evaluate the linearity of the assay. In the serum samples, the average detection rate of IFN-γ was 99.16%.
Intra-Assay Statistics
Ten replicates of each of four samples containing different IFN-γ concentrations were tested in one assay. Acceptable criteria: CV＜10%.
Inter-Assay Statistics
Four samples containing different concentrations of IFN-γ were tested in the independent assays. Acceptable criteria: CV<15%.
Recovery
IFN-γ was spiked into five human serum samples, and then analyzed. The average recovery of IFN-γ for serum samples is 95.92%.
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Interferon gamma
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