Human IFN-gamma ELISA

RayBio® Human IFN-gamma ELISA Kit for cell culture supernatants, plasma, serum, and lysate samples.

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Old price: $723.60
$578.88  
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ELH-IFNg-2
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Assay Format

Detection Range:
15 pg/ml - 15000 pg/ml
Recommended Dilution (serum & plasma):
2 fold
Sensitivity:
15 pg/ml

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Specificity:
This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
Number of Targets Detected:
1
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Cell Lysates
  • Plasma
  • Serum
  • Tissue Lysates
Design Principle:
  • Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
  • Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Protein Information

Accession Number:
  • P01579
Gene ID:
  • 3458
Gene Symbols:
  • IFNG
Protein Name & Synonyms:
Interferon gamma (IFN-gamma) (Immune interferon)
Target Species:
  • Human

Target Background

Interferon-gamma (IFN-gamma) is a pleiotropic cytokine with antiproliferative, pro-apoptotic and antitumor activity. It is a homodimer formed by the association of two 17 kDa polypeptides, that bind in an antiparallel manner to form a mature 50 kDa molecule. This symmetry allows for a single molecule to bind two receptors amplifying downstream signaling. IFN-gamma is secreted by CD4 T helper type 1 cells and CD8 cytotoxic cells, γδ T cells, NK cells and to a lesser extent by NK T cells, B cells and APC cells. Its expression is induced by cytokines such as IL-12, IL-15, IL-18 and type I IFN. The effects of IFN-gamma are mediated through the activation of the JAK/STAT pathway which modulates the transcription of many genes. IFN-gamma signaling plays a role in host defense, immune surveillance, establishing adaptive immunity and in regulating inflammation, apoptosis and the cell cycle.

Product Features

Additional Notes
  • 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

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components
  • 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
Other Materials Required
  • 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
Protocol Outline
  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.

Storage/Stability

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.
  1. Zhao J., Lu D., Moya S., et al. Bispecific T-cell engager (BiTE) immunotherapy of ovarian cancer based on MIL-88A MOF/MC gene delivery system. Applied Materials Today. Volume 20, September 2020, 100701
    Species: Human
    Sample type: Conditioned Media (T cells stimulatied with anti-CD3 and anti-Epcam BITE)
  2. Kundu-Raychaudhuri S., et al. Kv1.3 in psoriatic disease: PAP-1, a small molecule inhibitor of Kv1.3 is effective in the SCID mouse psoriasis e Xenograft model. Journal of Autoimmunity Aug 2014. epub ahead of print. http://dx.doi.org/10.1016/j.jaut.2014.07.003
    Species: Human
    Sample type: Conditioned Media (CD3 activated T cells with CD3/28)
  3. Fermoselle C., Rabinovich R., Ausin P., et al. Does oxidative stress modulate limb muscle atrophy in severe COPD patients? Eur Respir J. 2012 Oct;40(4):851-62.
    Species: Human
    Sample type: Tissue Lysate (Quadricep Biopsies from COPD patients)
  4. Liu Y., Liao J., Zhao M., et al. Increased expression of TLR2 in CD4(+) T cells from SLE patients enhances immune reactivity and promotes IL-17 expression through histone modifications. Eur J Immunol. 2015 Sep;45(9):2683-93. doi: 10.1002/eji.201445219.
    Species: Human
    Sample type: Conditioned Media (CD4 T cells with a TLR2 gene expression)
  5. Khalid M., Yu H., Sauter D., et al. Efficient Nef-mediated downmodulation of TCR-CD3 and CD28 is associated with high CD4+ T cell counts in viremic HIV-2 infection. J Virol. 2012 May;86(9):4906-20. doi: 10.1128/JVI.06856-11
    Species: Human
    Sample type: Conditioned Media (Human PBMCs infected with HIV constructs)
  6. Sambataro M, Sambado L., Trevisiol E., et al. Proinsulin-expressing dendritic cells in type 2 neuropathic diabetic patients with and without foot lesions. FASEB J. 2018 Feb 12:fj201701279RR. doi: 10.1096/fj.201701279RR
    Species: Human
    Sample type: Plasma (Diabetic Patient Samples)
  7. Sambataro M, Sambado L, Trevisiol E, Cacciatore M, Furlan A, Stefani PM, Seganfreddo E, Durante E, Conte S, Della Bella S, Paccagnella A, Dei Tos AP. Proinsulin-expressing dendritic cells in type 2 neuropathic diabetic patients with and without foot lesions. FASEB J. 2018 Jul;32(7):3742-3751. doi: 10.1096/fj.201701279RR. Epub 2018 Feb 12. PMID: 29436863.
    Species: Human
    Sample type: Plasma (diabetes and NC patients)
  8. Tammali, Ravinder, et al. "Inhibition of aldose reductase prevents angiogenesis in vitro and in vivo." Angiogenesis 14.2 (2011): 209-221.
    Species: Human
    Sample type: Conditioned Media
  9. ABDEL?HAMID, Mahmoud F., et al. "Serum levels of interleukin?8, tumor necrosis factor?? and ??interferon in Egyptian psoriatic patients and correlation with disease severity." The Journal of dermatology 38.5 (2011): 442-446.
    Species: Human
    Sample type: Serum
  10. Yu H, Usmani SM, Borch A, et al. The efficiency of Vpx-mediated SAMHD1 antagonism does not correlate with the potency of viral control in HIV-2-infected individuals. Retrovirology. 2013;10:27.
    Species: Human
    Sample type: Conditioned Media (HIV-infected dendritic cells)
Well trusted material for my analysis

University of Puerto Rico
we used this kit to detect IFN gamma in iPSC-derived CNS cells. it works fine. it took about 4.5 hours to complete the assay which is reasonable.

Mayo Clinic
Plasma & Urine were run at 2-fold dilution. Urine samples were BLQ. Plasma samples had very low concentration. Our samples and too low of a concentration to be measured by this assay.

University of Michigan
This ELISA kit was used for quantification of IFN-gamma in the supernatants of SEB-stimulated peripheral blood mononuclear cells in combination with conventional chemotherapeutics. The variability of the technical replicates was low and the data obtained very repetitive.

KYNAN Pharma
This human IFNg ELISA kit works very well. Steps included in the manufacturer's protocol were easy to follow and all the necessary materials were provided. The snap wells come in handy when running multiple experiments in small batches for optimization of the sample. We ran human plasma samples and the standards ran evenly and the detection concentration fell within the provided range which is rare for quantifying IFNg levels in human samples.

University of Illinois at Chicago
This is the first time I've run an Interferon-g ELISA and I'm very happy with the results. I typically use the ELISpot platform and I'm happy that the results with this kit are far superior- very little background and fairly sensitive with the detection. I may try the newer version (with the qRT-PCR) readout for subsequent experiments to compare between kits.

Mayo Clinic
This human IFNg ELISA kit is good. Steps were clear and all necessary materials were provided. Since we ran multiple smaller experiments, snap wells were really handy. We used BAL from mice lungs and though the standard ran evenly, samples showed low IFNg levels.

University of Pittsburgh
The human IFNg ELISA kit is easy to follow and reproducible. We used the kit with IFNg-secreting human cell lines following the manufacturer's protocol after the cells were treated with LPS. The supernatant was used to detect the IFNg. The kit worked perfectly when detecting high content of IFNg. However, when the IFNg content is low in the samples, more repeats may be needed as the sensitivity or accuracy of the kit is becoming low, too. Overall, the kit works well in detecting IFNg.
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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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