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ELISpot MAX™ Deluxe Set Human IFN-γ

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Regulatory Status
RUO
Other Names
Interferon gamma, IFN-gamma
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ELISpot-MAX_Human-IFN-gamma_121923
Human peripheral blood mononuclear cells (PBMC) at 104 cells /well were incubated for 20 hours in the absence or presence of 1X Cell Activator without Brefeldin A (Cat. No. 423301), 1X Cell Activator with Brefeldin A (Cat. No. 423303), or Ultra-LEAF™ purified anti-human CD3 (Cat. No. 300331). BioLegend’s Cell Activator cocktail contains PMA and ionomycin at the optimized concentration. Brefeldin A functions to inhibit intracellular protein transport. As a result, it inhibits secretion of cytokines such as IFN-γ. The number of cells secreting human IFN-γ was analyzed by ELISpot.
  • ELISpot-MAX_Human-IFN-gamma_121923
    Human peripheral blood mononuclear cells (PBMC) at 104 cells /well were incubated for 20 hours in the absence or presence of 1X Cell Activator without Brefeldin A (Cat. No. 423301), 1X Cell Activator with Brefeldin A (Cat. No. 423303), or Ultra-LEAF™ purified anti-human CD3 (Cat. No. 300331). BioLegend’s Cell Activator cocktail contains PMA and ionomycin at the optimized concentration. Brefeldin A functions to inhibit intracellular protein transport. As a result, it inhibits secretion of cytokines such as IFN-γ. The number of cells secreting human IFN-γ was analyzed by ELISpot.
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430124 5 plates 492€
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Description

Human IFN-γ (Interferon-gamma) is a multifunctional cytokine which is the sole member of the class II Interferon. Although a variety of immune cells can express IFN-γ, most of IFN-γ is produced by T-cells and NK cells. It has critical roles in many processes including, but not limited to, anti-viral response, regulating Th1/Th2 balance, enhancing antigen presentation, and controlling cellular proliferation and apoptosis. IFN-γ can bind to 2 receptors: IFNGR1 and IFNGR2, which transduce the signal downstream through the JAK-STAT pathway. Because of its multifunction nature, dysregulation of IFN-γ expression and activity has been implicated in many diseases such as autoimmune and cancer. 

BioLegend’s ELISpot MAX™ Deluxe Set contains pre-optimized essential reagents and additional buffers and solution for Enzyme-Linked Immunosorbent Spot assay. This kit is designed for detection of human IFN-γ secreting cells. A monoclonal antibody specific for human IFN-γ is coated on ethanol-activated 96-well PVDF membrane plate as the capture. Stimulated and/or unstimulated cells are pipetted directly into the wells, and then incubated at 37°C for a specific period of time. During this period, capture antibody binds to human IFN-γ from the secreting cells in the vicinity. Then, cells are removed, and a biotinylated anti-human IFN-γ detection antibody is added, followed by Avidin-HRP. Subsequently, TMB membrane substrate is added. The TMB membrane substrate reacts with HRP to form a blue-black precipitate at the sites of human IFN-γ secreting cells. The end result is visible spots, which correspond to footprints of IFN-γ secreting cells on the membrane surface.

Product Details
Technical data sheet

Kit Contents

Kit Contents
  • Human IFN-γ ELISpot MAX™ Capture Antibody (200X)
  • Human IFN-γ ELISpot MAX™ Detection Antibody (200X)
  • Human IFN-γ Standard (used as positive control)
  • Avidin-HRP (1000X)
  • TMB Membrane Substrate
  • Coating Buffer A (5X)
  • Assay Diluent B (5X)

Product Details

Verified Reactivity
Human
Application

ELISpot
Materials Not Included
  • PVDF-bottom plate: Millipore MultiScreenHTS IP Filter Plate (Cat. No. MSIPS4W10) is recommended.
  • Wash Buffer: BioLegend Cat. No. 421601 is recommended, or PBS + 0.05% Tween-20  
  • Plate Sealers: BioLegend Cat. No. 423601 is recommended 
  • 0.22 μm filters
  • 35% Ethanol
  • Sterile PBS (Phosphate-Buffered Saline): 8.0 g NaCl, 1.16 g Na2HPO4, 0.2 g KH2PO4, 0.2 g KCl, add deionized water to 1 L; pH to 7.4, 0.22 μm filtered
  • Sterile cell culture media
  • 37°C CO2 incubator
  • Deionized (DI) water 
  • Adjustable pipettes to measure volumes ranging from 2 μL to 1 mL 
  • Wash bottle, manifold dispenser, or automated microplate washer 
  • Tubes for dilution
  • Timer 
  • Stereomicroscope or an automated ELISpot reader

Antigen Details

Cell Type
NK cells, T cells
Biology Area
Immuno-Oncology, Immunology, Innate Immunity, Neuroscience, Signal Transduction
Molecular Family
Cytokines/Chemokines, Innate Immune Signaling, Tumor Suppressors
Gene ID
3458 View all products for this Gene ID
UniProt
View information about IFN-gamma on UniProt.org
Go To Top Version: 1    Revision Date: 12.19.2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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