Human IFN-γ(Interferon Gamma) ELISA Kit

    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience

      Catalog number:E-EL-H0108

      Synonyms:IFNG, IFG, IFI, Type II Interferon

      Size:
      • 96T
      • 24T
      Qty:
      - +
      Price: $495

      Reactivity: Human

      Detection Range: 15.63~1000 pg/mL

      Sensitivity: 9.38 pg/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human IFN-γ concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human IFN-γ. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human IFN-γ and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human IFN-γ, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human IFN-γ. You can calculate the concentration of Human IFN-γ in the samples by comparing the OD of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 3.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 15.63—1000 pg/mL
      Sensitivity 9.38 pg/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human IFN-γ in samples. No significant cross-reactivity or interference between Human IFN-γ and analogues was observed.

      Typical data

      As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

      Concentration
      (pg/mL)
      O.D Average Corrected
      1000 2.475
      2.517
      2.496 2.438
      500 1.552
      1.602
      1.577 1.519
      250 0.963
      0.953
      0.958 0.9
      125 0.463
      0.487
      0.475 0.417
      62.5 0.251
      0.241
      0.246 0.188
      31.25 0.171
      0.145
      0.158 0.1
      15.63 0.107
      0.113
      0.11 0.052
      0 0.048
      0.068
      0.058 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human IFN-γ were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human IFN-γ were tested on 3 different plates, 20 replicates in each plate.

      Intra-assay Precision Inter-assay Precision
      Sample 1 2 3 1 2 3
      n 20 20 20 20 20 20
      Mean (pg/mL) 51.90 98.00 447.20 55.80 100.10 476.70
      Standard deviation 3.20 4.80 16.10 2.80 5.40 24.30
      C V (%) 6.17 4.90 3.60 5.02 5.39 5.10

      Recovery

      The recovery of Human IFN-γ spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

      Sample Type Range (%) Average Recovery (%)
      Serum (n=5) 96-107 101
      EDTA plasma (n=5) 89-104 96
      Cell culture media (n=5) 86-97 91

      Linearity

      Samples were spiked with high concentrations of Human IFN-γ and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

      Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
      1:2 Range (%) 99-114 92-105 89-105
      Average (%) 105 98 96
      1:4 Range (%) 98-110 86-99 91-105
      Average (%) 103 92 97
      1:8 Range (%) 96-110 82-98 98-114
      Average (%) 102 89 104
      1:16 Range (%) 97-111 85-99 95-109
      Average (%) 105 90 102

      Kit components & Storage

      An unopened kit can be stored at 2-8°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

      Item Specifications Storage
      Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20°C, 6 months
      Reference Standard 2 vials
      Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
      Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20°C(shading light), 6 months
      Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months
      Biotinylated Detection Ab Diluent 1 vial, 14 mL
      HRP Conjugate Diluent 1 vial, 14 mL
      Concentrated Wash Buffer (25×) 1 vial, 30 mL
      Substrate Reagent 1 vial, 10 mL 2-8°C(shading light)
      Stop Solution 1 vial, 10 mL 2-8°C
      Plate Sealer 5 pieces
      Product Description 1 copy
      Certificate of Analysis 1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

      Other supplies required

      • Microplate reader with 450 nm wavelength filter
      • High-precision transfer pipette, EP tubes and disposable pipette tips
      • Incubator capable of maintaining 37°C
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      elisa assay procedure 1

      1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C

      elisa assay procedure 2

      2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C

      elisa assay procedure 3

      3. Aspirate and wash the plate for 3 times

      elisa assay procedure 4

      4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

      elisa assay procedure 5

      5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

      elisa assay procedure 6

      6. Add 50μL Stop Solution

      elisa assay procedure 7

      7. Read the plate at 450nm immediately. Calculation of the results

      Citations

      1. Publication: Xia G, Zheng X, Yao X, et al. Expression Of Programmed Cell Death-1 And Its Ligand B7 Homolog 1 In Peripheral Blood Lymphocytes From Patients With Peripartum Cardiomyopathy[J]. Clinical Cardiology, 2016.
        Species: Human
        Sample Type: Serum
      2. Publication: Li W, Dong H, Huang Y, et al. Clonorchis Sinensis Co-Infection Could Affect The Disease State And Treatment Response Of Hbv Patients[J]. Plos Neglected Tropical Diseases, 2016.
        Species: Human
        Sample Type: Culture supernatants and Serum
      3. Publication: Xia G, Xin S, Zheng X, et al. Decreased Expression Of Programmed Death 1 On Peripheral Blood Lymphocytes Disrupts Immune Homeostasis In Peripartum Cardiomyopathy[J]. International Journal of Cardiology, 2016, 223: 842-847.
        Species: Human
        Sample Type: Serum
      4. Publication: Wu R X, Yu Y, Yin Y, et al. Platelet Lysate Supports The In Vitro Expansion Of Human Periodontal Ligament Stem Cells For Cytotherapeutic Use[J]. Journal of Tissue Engineering And Regenerative Medicine, 2016.
        Species: Human
        Sample Type: Culture supernatant
      5. Publication: He W, Ren Y, Wang X, et al. C Reactive Protein And Enzymatically Modified LDL Cooperatively Promote Dendritic Cell-Mediated T Cell Activation[J]. Cardiovascular Pathology, 2017, 29: 1-6.
        Species: Human
        Sample Type: Culture supernatant
      6. Publication: Wang Z Y, Jiang M J, Wu M H, et al. Inflammatory Characteristics In Different Types Of Nonallergic Rhinitis[J]. International Journal of Clinical and Experimental Pathology, 2017, 10(3): 2887-2894.
        Species: Human
        Sample Type: Culture supernatant
      7. Publication: Wang C M, Zhang T, Wang Y H, et al. The Shortening Telomere Length Of T Lymphocytes Maybe Associated With Hyper‑Function In Servere Aplastic Anemia[J]. Molecular Medicine Reports, 2018, 17: 1015-1021.
        Species: Human
      8. Publication: Wu R X, Yu Y, Yin Y, et al. Platelet Lysate Supports The In Vitro Expansion Of Human Periodontal Ligament Stem Cells For Cytotherapeutic Use[J]. Journal of Tissue Engineering And Regenerative Medicine, 2017, 11(8): 2261-2275.
        Species: Human
        Sample Type: Culture supernatant
      9. Publication: Gao J, Wei L T, Liu X H, et al. Association Between IFN-γ Gene Polymorphisms And IgA Nephropathy In A Chinese Han Population[J]. Kidney & Blood Pressure Research, 2017, 42(1): 136-144.
        Species: Human
        Sample Type: Serum
      10. Publication: Giusti I, Di Francesco M D, D'Ascenzo S, et al. Leukocyte Depletion Does Not Affect The In Vitro Healing Ability Of Platelet Rich Plasma[J]. Experimental and Therapeutic Medicine, 2018, 15(4): 4029-4038.
        Species: Human
      11. Publication: Wang L, Yang X, Li D, et al. The elevated glutaminolysis of bladder cancer and T cells in a simulated tumor microenvironment contributes to the up-regulation of PD-L1 expression by interferon-γ. Onco Targets Ther. 2018;11:7229-7243. Published 2018 Oct 18. doi:10.2147/OTT.S180505
        Species: Human
        Sample Type: Cell culture supernatant
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