IFNG ELISA Kit (Interferon gamma)

Details for Product IFNG ELISA Kit No. ABIN625130, Supplier: Log in to see
  • IFG
  • IFI
  • IFN-g
  • Ifg
  • IFNG2
  • IFN-gamma
  • IFN-G
  • IFNG
  • IFNgamma
  • TCRalpha
  • INF-G
  • ifng
  • interferon gamma
  • interferon, gamma 1-2
  • IFNG
  • Ifng
  • ifng1-2
Mouse (Murine)
Kits with alternative reactivity to:
Method Type
Sandwich ELISA
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Supplier Product No.
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Purpose Mouse IFN-gamma ELISA Kit for cell and tissue lysate samples.
Sample Type Tissue Lysate, Cell Lysate
Analytical Method Quantitative
Detection Method Colorimetric
Specificity The antibody pair provided in this kit recognizes mouse IFN-gamma.
Sensitivity 10 pg/mL
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Material not included
  • Distilled or deionized water
  • Precision pipettes to deliver 2 μL to 1 μL volumes
  • Adjustable 1-25 μL pipettes for reagent preparation
  • 100 μL and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
  • Cell lysate buffer
Plasmids, Primers & others Plasmids, Primers & others IFNG products on genomics-online (e.g. as negative or positive controls)
Alternative Name IFN-gamma (IFNG ELISA Kit Abstract)
Background IFN-gamma is produced mainly by T-cells and natural killer cells activated by antigens, mitogens, or alloantigens. It is produced by lymphocytes expressing the surface antigens CD4 and CD8. Mouse IFN-gamma is a polypeptide of 136 amino acids, containing four exons and three introns. It plays an important role in the immune IFN-gamma response. IFN-gamma is a modulator of T-cell growth and functional differentiation. It is a growth-promoting factor for T-lymphocytes and potentiates the response of these cells to mitogens or growth factors. The Mouse IFN-gamma ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse IFN-gamma in cell lysate and tissue lysate. This assay employs an antibody specific for mouse IFN-gamma coated on a 96-well plate. Standards and samples are pipetted into the wells and IFN-gamma present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-mouse IFN-gamma antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IFN-gamma bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.
Gene ID 15978
UniProt P01580
Pathways Interferon-gamma Pathway, Cellular Response to Molecule of Bacterial Origin, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, ER-Nucleus Signaling, Regulation of Carbohydrate Metabolic Process, Protein targeting to Nucleus, Autophagy
Sample Volume 100 μL
Plate Pre-coated
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 μL of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4 °C.
  4. Add 100 μL of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 μL of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 μL of Stop Solution to each well.
  11. Read at 450 nm immediately.
Reagent Preparation
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
    2. Sample dilution: Tissue lysate and cell lysate sample should be diluted at least 5-fold with Sample Diluent Buffer.
    3. Sample Diluent Buffer (Item D) and Assay Diluent (Item E) should be diluted 5-fold with deionized or distilled water before use.
    4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL 1x Sample Diluent Buffer (Item D should be diluted 5-fold with deionized or distilled water before use) into Item C vial to prepare a 20 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 50 µL IFN-gamma standard from the vial of Item C, into a tube with 450 µL 1x Sample Diluent Buffer to prepare a 2,000 pg/mL stock standard solution. Pipette 400 µL 1x Sample Diluent Buffer into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. 1x Sample Diluent Buffer serves as the zero standard (0 pg/mL). 200 µL 200myl 200 µL 200 µL 200 µL 200 µL 50 µL standard + 450 µL 2,000 666.7 222.2 74.07 24.67 8.23 2.74 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
    5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
    6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 120-fold with 1x Assay Diluent and used in step 4 of Part VI Assay Procedure.
    7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 440-fold with 1x Assay Diluent. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 25 µL of HRP-Streptavidin concentrate into a tube with 11 ml 1x Assay Diluent to prepare a 440-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
    8. Cell lysate buffer should be diluted 2-fold with deionized or distilled water (for cell lysate and tissue lysate).
Assay Procedure
  1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
    2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking. We recommend using 50-500 myg/mL of total protein for lysate sample. The amount of sample used depends on the abundance of target protein. More of the sample can be used if signals are too weak. If signals are too strong, the sample can be diluted further.
    3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
    4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking.
    5. Discard the solution. Repeat the wash as in step
    6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
    7. Discard the solution. Repeat the wash as in step
    8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
    9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.
Calculation of Results

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Sample Diluent Buffer Mouse IFN-gamma concentration (pg/mL) 1 10 100 1000 10000 O D =4 50 n m 0.01 0.1 1 10
Sensitivity: The minimum detectable dose of IFN-gamma is typically less than 10 pg/mL.
Recovery: Recovery was determined by spiking various levels of mouse IFN-gamma into tissue lysate and cell lysate. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Tissue lysate 93.47 83-103 Cell lysate 91.95 84-104
Linearity: Sample Type Tissue Cell Lysate lysate 1:2 Average % of 91 92 Expected Range ( %) 80-103 84-104 1:4 Average % of 97 94 Expected Range ( %) 84-104 85-103
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Assay Precision Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
Restrictions For Research Use only
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Expiry Date 6 months
Supplier Images
ELISA image for Interferon gamma (IFNG) ELISA Kit (ABIN625130) Interferon gamma (IFNG) ELISA Kit
Product cited in: Wu, You, Ma, Li, Yuan, Li, Ye, Liu, Yao, Chen, Lai, Yang: "Role of transient receptor potential ion channels and evoked levels of neuropeptides in a formaldehyde-induced model of asthma in BALB/c mice." in: PLoS ONE, Vol. 8, Issue 5, pp. e62827, 2013

Wu, You, Ma, Li, Yuan, Li, Ye, Liu, Yao, Chen, Lai, Yang: "Role of transient receptor potential ion channels and evoked levels of neuropeptides in a formaldehyde-induced model of asthma in BALB/c mice." in: PLoS ONE, Vol. 8, Issue 5, pp. e62827, 2013 (PubMed).

Dalton, Pitts-Meek, Keshav, Figari, Bradley, Stewart: "Multiple defects of immune cell function in mice with disrupted interferon-gamma genes." in: Science (New York, N.Y.), Vol. 259, Issue 5102, pp. 1739-42, 1993 (PubMed).

Gu, Sarvetnick: "Epithelial cell proliferation and islet neogenesis in IFN-g transgenic mice." in: Development (Cambridge, England), Vol. 118, Issue 1, pp. 33-46, 1993 (PubMed).

Cooper, Dalton, Stewart, Griffin, Russell, Orme: "Disseminated tuberculosis in interferon gamma gene-disrupted mice." in: The Journal of experimental medicine, Vol. 178, Issue 6, pp. 2243-7, 1993 (PubMed).

Naylor, Gray, Lalley: "Mouse immune interferon (IFN-gamma) gene is on chromosome 10." in: Somatic cell and molecular genetics, Vol. 10, Issue 5, pp. 531-4, 1984 (PubMed).

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