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Human IFN-γ / Perforin Dual ELISpot Set

Product Specifications

Catalogue N°
874.070.001 - 1 x 96 Discovery (plate not included)
874.070.001P - 1 x 96 Discovery (non-sterile plate)
874.070.001S - 1 x 96 Discovery (sterile plate)
874.070.005 - 5 x 96 (plates not included)
874.070.005P - 5 x 96 (non-sterile plates)
874.070.005S - 5 x 96 (sterile plates)
874.070.010 - 10 x 96 (plates not included)
874.070.010P - 10 x 96 (non-sterile plates)
874.070.010S - 10 x 96 (sterile plates)
874.070.015 - 15 x 96 (plates not included)
874.070.015P - 15 x 96 (non-sterile plates)
874.070.015S - 15 x 96 (sterile plates)
874.070.020 - 20 x 96 (plates not included)
874.070.020P - 20 x 96 (non-sterile plates)
874.070.020S - 20 x 96 (sterile plates)
Target species
Human
Specificity
Recognizes natural human IFN-g and Perforin
Incubation
3h30 after cell stimulation
Cross Reaction
No cross reactivity with other human cytokines. Cross reactivity with simian IFN-g
Kit Content
Diaclone Dual ELISpot Sets include capture and detection antibodies for both of the targeted cytokines, Streptavidin-Alkaline Phosphatase conjugated, anti-FITC HRP conjugated mAb, BSA, BCIP/NTB and AEC blocking reagent.

BACKGROUND

IFNg

IFNg, also called Type II interferon, is a homodimeric glycoprotein containing approximately 21 to 24 kD subunits. The human IFNg gene, situated on chromosome 12, contains three introns; the four exons code for a polypeptide of 166 amino acids, 20 of which constitute the signal peptide. In contrast to IFNa and IFNb synthesis, which can occur in any cell, production of IFNg is a function of T cells and NK cells. All IFNg inducers activate T cells either in a polyclonal (mitogens or antibodies) or in a clonally restricted, antigen-specific, manner. IFNg is produced during infection by T cells of the cytotoxic/suppressor phenotype (CD8) and by a subtype of helper T cells, the Th1 cells. Th1 cells secrete IL-2, IL-3, TNFa and IFNg, whereas Th2 cells main produce IL-3, IL-4, IL-5, and IL-10, but little or no IFNg. IFNg preferentially inhibits the proliferation of Th2 but not Th1 cells, indicating that the presence of IFNg during an immune response will result in the preferential proliferation of Th1 cells .

Type II IFN or IFNg is a lymphokine that displays no molecular homology with type I IFN, but shares some important biologic activities. Specifically, IFNg induces an anti-viral state and is anti-proliferative. In addition, IFNg has several properties related to immunoregulation.

1) IFNg is a potent activator of mononuclear phagocytes, e.g. IFNg stimulates the expression of Mac-1, augments endocytosis and phagocytosis by monocytes, and activates macrophages to kill tumor cells by releasing reactive oxygen intermediates and TNFa.

2) IFNg induces or augments the expression of MHC antigens on macrophages, T and B cells and some tumor cell lines.

3) On T and B cells IFNg promotes differentiation. It enhances proliferation of activated B cells and can act synergistically with IL-2 to increase immunoglobulin light-chain synthesis . IFNg is one of the natural B-cell differentiation factors.

4) Finally, IFNg activates neutrophils, NK cells and vascular endothelial cells.

The role of IFNg as a disease marker has been demonstrated for a number of different pathological situations:

  • IFNg is produced during viral infections. IFNg is a diagnostic tool for distinguishing tuberculous from other non-tuberculous ascites . IFNg values in pleural fluid are significantly higher in tuberculous pleuritis patients than those in non-tuberculous pleuritis patients, with a sensitivity and a specificity of 100%. Therapy-induced (treatment with thalidomide) alleviation of clinical symptoms of erythema nodosum leprosum correlates with IFNg and TNFa levels. Tuberculoid leprosy patients show increased lymphocyte proliferation and IFNg production in response to stimulation with Mycobacterium leprae as compared to lepromatous leprosy patients and healthy individuals.
  • Autoimmune diseases: Accurate measurements of cellular production of cytokines, e.g. IFNg is important in the design and monitoring of immunotherapy of multiple sclerosis.
  • Transplant rejection: Intragraft IFNg mRNA expression occurs in active acute transplant rejection preceding clinical transplant rejection, thus offering an early diagnostic tool for detection of transplant rejection.
  • IFNg production by isolated lymphocytes is not detectable in patients with cow's milk allergy as compared to control individuals. Infants who develop atopy produce significantly less IFNg at birth compared to infants who do not develop atopy.
  • Peripheral blood lymphomononuclear cells from newly diagnosed type I diabetes produce significantly less IFNg in comparison to controls and long standing diabetes.

 

Perforin

Perforin is a 70kDa protein containing 534 amino acid.

Perforin is involved in the killing function by CTLs and NKs. Upon degranulation, perforin inserts itself into the target cell's plasma membrane, forming a pore. It leads to osmotic lysis of the target cells and subsequently enables granzymes to enter the target cells and activate apoptosis, the cell death program.

Perforin has structural and functional similarities to complement component 6, 7, 8 and 9 (C6, C7, C8, C9). Like C9, this protein creates transmembrane tubules and is capable of lysing non-specifically a variety of target cells. This protein is one of the main cytolytic proteins of cytolytic granules, and it is known to be a key effector molecule for T-cell- and natural killer-cell-mediated cytolysis.

Its role in the immune response against tumors and virus infection has been demonstrated.

Version 6 - 07.19

For research use only

Documents

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Formats Available

874.070.001 1 x 96 Discovery (plate not included)
874.070.001P 1 x 96 Discovery (non-sterile plate)
874.070.001S 1 x 96 Discovery (sterile plate)
874.070.005 5 x 96 (plates not included)
874.070.005P 5 x 96 (non-sterile plates)
874.070.005S 5 x 96 (sterile plates)
874.070.010 10 x 96 (plates not included)
874.070.010P 10 x 96 (non-sterile plates)
874.070.010S 10 x 96 (sterile plates)
874.070.015 15 x 96 (plates not included)
874.070.015P 15 x 96 (non-sterile plates)
874.070.015S 15 x 96 (sterile plates)
874.070.020 20 x 96 (plates not included)
874.070.020P 20 x 96 (non-sterile plates)
874.070.020S 20 x 96 (sterile plates)

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