FluoroSpot Plus: Monkey IFN-γ/IL-17A

FluoroSpot Plus: Monkey IFN‑γ/IL‑17A

FSP-2125-2

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Components

PlatePre-coated FluoroSpot plate (mAbs MT126L and MT241)
Detection mAbsAnti-IFN-γ mAb (7-B6-1), BAM
Anti-IL-17A mAb (MT504), biotin
Fluorophore conjugatesAnti-BAM mAb, 490
SA-550
Buffer/SolutionFluoroSpot enhancer
StimulusAnti-CD3 mAb (CD3-1)
Co-stimulusAnti-CD28 mAb (CD28-A)

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Product specifications

Intended use

This FluoroSpot Plus kit is intended for the enumeration of cells secreting monkey IFN-γ and IL-17A using the FluoroSpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Recommendation

Product details

ProductFluoroSpot Plus: Monkey IFN-γ/IL-17A
ApplicationFluoroSpot
AnalyteIFN-γ, IL-17A
ReactivityNon-human primates
Specificity

This kit is based on a matched pair of mAbs enabling specific detection of IFN-γ and IL-17A from non-human primates (NHP).

Cross-reactivity

The mAbs react with human IFN-γ and IL-17A, and cross-react with IFN-γ and IL-17A from NHP. For more information please check the Cross-reactivity guide section below.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store antibodies, fluorophore conjugates, and FluoroSpot enhancer at 4-8 °C upon receipt. Plates may be kept at room temperature.

Shelf lifeAt least 12 months from date of receipt.
PBMC from cynomolgus macaque (415,000 cells/well) were incubated overnight in the presence of PHA and the number of cells secreting IFN-γ and/or IL-17A were analyzed by FluoroSpot. Double-secreting cells were determined as spots having the same center point in an image overlay of LED490 (IFN-γ) and LED550 (IL-17A) images. Spot analysis was made with Mabtech IRIS.

The systems reactive with NHPs are either based on cross-reactive human kits or specifically developed monkey kits (NHP).

Cross-reactivity verification tests have been performed in ELISpot and/or ELISA by Mabtech and/or by others. Evaluations of assays with less solid evidence of cross-reactivity are shown as symbols within parentheses.

 Rhesus MacaqueCynomolgus MacaquePig-tailed MacaqueBaboon*Sooty MangabeyAfrican Green MonkeyNight Monkey*Common MarmosetSquirrel Monkey
ASSAYOld worldNew world
ApoA1      
ApoB       
ApoE (NHP)       
ApoE       
ApoH       
CCL2 (MCP-1)       
CCL4 (MIP-1β) ( ) ( )( )( )( )
CCL22 (MDC) ( ) ( )( )( )( )
CD25       
GM-CSF( )( )   
Granzyme A        
Granzyme B (NHP)       
Granzyme B     
IFN-α2        
IFN-α pan   ( )   
IFN-γ (NHP)
IFN-γ( )
IgA (NHP)       
IgA       
IgG       
IgM       
INS     
IL-1α        
IL-1β        
IL-2 (NHP)
IL-3       
IL-4( )( )( )( )
IL-5( )( )( ) 
IL-6( )( ) ( ) 
IL-8 (NHP)        
IL-8 (ELISA)        
IL-8 (ELISpot)        
IL-10 (NHP)      
IL-10       
IL-12/-23 (p40)      
IL-12 (p70)        
IL-13 (NHP)( )( )( )  
IL-13        
IL-17A (NHP)( )( )   
IL-17A     
IL-17F        
IL-17A/F        
IL-21      
IL-22        
IL-23     
IL-27        
IL-31        
IP-10       
Perforin( )   
TGF-β1 (latent)       
TNF-α (NHP)( )  ( )( )( )( )
          
SINGLE mAbs         
Cell stimulation         
CD3, mAb CD3-1       
CD3, mAb CD3-2       
CD28, mAb CD28-A       
Flow cytometry          
IFN-γ mAb 1-D1K( )( )( )( )( )( )
IL-2 mAb MT8G10( )( )( )( ) ( )( )( )
IL-4 mAb IL4-3( )       
IL-17A mAb MT504        
Perforin mAb Pf-344     
TNF-α mAb MT15B15       
Ig reagents         
IgG1 mAb MTG1218       
IgG1 mAb MT1939       
IgG2 mAb H6200       
IgG2 mAb MTG211E       
IgG3 mAb MTG34       
IgG4 mAb MTG42       
Western blot         
IFN-γ mAb MT111W( )( )( )( )( )( )( )( )( )
TNF-α mAb MT15B15( )( )( )  ( )( )( )( )

Symbol keys:


Cross-reactivity

Verified by Mabtech and/or by others. Tests performed in ELISpot and/or ELISA.

  • good
  • poor
  • no

Indications of Cross-reactivity

Based on reports from others or by analysis using recombinant proteins.

  • ( ) good
  • ( ) poor
  • ( ) no

Monkey kits

Specifically developed monkey kits are marked with (NHP).

Blank boxes

Represent cross-reactivities not evaluated.

* Comprises several species. Cross-reactivity may have to be verified on a species basis.

Tutorial, Published November 24, 2022

Cell stimulation in ELISpot and FluoroSpot
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IFN-γ

Analyte description

Interferon-γ (IFN-γ) is the only type II interferon. This proinflammatory cytokine is secreted by activated T cells and NK cells. It activates macrophages and endothelial cells and regulates immune responses by affecting APCs, T cells, and B cells. Production of IFN-γ by helper T cells and cytotoxic T cells is a hallmark of the Th1-type phenotype. Thus, high-level production of IFN-γ is typically associated with effective host defense against intracellular pathogens.

Alternative namesInterferon-γ, Interferon-gamma, IFN-γ, IFN-gamma, IFN-g, IFNg, IFG, IFI, IMD69
Cell typeT cell, Tc, Th1, NK cell
Gene ID3458

IL-17A

Analyte description

Interleukin 17A (IL-17A) is a potent proinflammatory cytokine produced by activated Th17 (T helper 17) cells and certain cells belonging to the innate immune system. In mice, IL-17 has also been shown to be produced by activated CD8 T cells and γδ T cells. Th17 cells play an important role in autoimmune diseases and protection against bacteria and fungi. IL-17A acts on a broad range of cell types to induce the expression of cytokines, chemokines, and metalloproteinases. As a result, secretion of IL-17A promotes inflammatory responses, which leads to the recruitment of neutrophils, enhancement of antibody production, and activation of T cells. Increased expression of IL-17A is seen in autoimmune diseases such as multiple sclerosis and rheumatoid arthritis. It is also associated with asthma, psoriasis, cancer, and transplant rejection.

Alternative namesInterleukin 17A, IL-17A, IL17A, CTLA-8, CTLA8, IL-17, IL17, ILA17
Cell typeTh17
Gene ID3605