Anti-bovine IFN-γ mAb (MT17.1), unconjugated

Anti-bovine IFN-γ mAb (MT17.1), unconjugated

3119-3-250

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Product specifications

Intended use

This monoclonal antibody enables specific detection of bovine IFN-γ in immunoassays such as ELISpot, FluoroSpot, and ELISA.

Serum/Plasma samples

Recommendation

MT17.1 is recommended as capture mAb in ELISpot, FluoroSpot, and ELISA in combination with detection mAb MT307 (product code 3119-6).

Product details

ProductAnti-bovine IFN-γ mAb (MT17.1), unconjugated
ApplicationELISpot, FluoroSpot, ELISA
AnalyteIFN-γ
AntibodyMT17.1
ConjugateUnconjugated
ClonalityMonoclonal
ImmunogenRecombinant bovine IFN-γ
HostMouse
IsotypeIgG1
ReactivityCow, Sheep
Specificity

Native and recombinant bovine (cow) IFN-γ

Cross-reactivity

Cross-reacts with IFN-γ from sheep. For more information, please check the Veterinary cross-reactivity guide.

Purification

Purified from in vitro cultures by protein G affinity chromatography.

Concentration0.5 mg/ml
Supplied in

PBS with 0.02% sodium azide. Sterile-filtered (0.2 µm).

Contents

Monoclonal antibody MT17.1, unconjugated. Supplied at 0.5 mg/ml in PBS with 0.02% sodium azide.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store product at 4-8°C or frozen at -20°C or below. Avoid repeated freezing/thawing.

Shelf lifeAt least 18 months from date of receipt.

Find out which analyte combinations we have evaluated in T cell FluoroSpot and which combinations are affected by capture effects or not. 

AnalytesGM-CSFGzAGzBIFN-aIFN-γIgAIgGIgMIL-1αIL-1βIL-2IL-3IL-4IL-5IL-6IL-8IL-10IL-12 (p70)IL-12/-23IL-13IL-17AIL-21IL-22IL-27IL-29 (IFN-λ1)TNF-α
GM-CSF         *             
GzA                        
GzB   *     *    *        *
IFN-a                         
IFN-γ *       *     
IgA                        
IgG                        
IgM                        
IL-1α                      
IL-1β*                     
IL-2  * *           *   
IL-3                       
IL-4                        
IL-5            *   *   
IL-6                     
IL-8                          
IL-10  *       *     *    
IL-12 (p70)                          
IL-12/-23                         
IL-13         *         *   
IL-17A             *    *   
IL-21                         
IL-22           *     **     
IL-27                         
IL-29 (IFN-λ1)                         
TNF-α *                 

 

Symbol keys:

Verified by Mabtech, mainly with human samples

  •    works well
  • * recommended to include anti-CD28
  •    major capture effects observed
  • Blank boxes indicate combinations not evaluated

What is a capture effect?

When capture antibodies with different specificities are coated together, the capture of one cytokine may affect the secretion of other cytokines. This is usually more pronounced when studying T cell responses with polyclonal stimuli compared to antigen-specific responses. Capture effects are seldom a problem and can often be counteracted by the addition of an anti-CD28 antibody.

Depiction of capture effects observed in FluoroSpot

(1) IL-2 secreted by the activated T cell is captured by coated anti-IL-2 capture antibodies. (2) As a result, IL-2-stimulation of the T cell itself (autocrine stimulation) as well as nearby T cells (paracrine stimulation) is impaired, ultimately leading to (3) decreased IFN-γ secretion.

Co-stimulation with anti-CD28

Anti-CD28 mAb provides a co-stimulatory signal to antigen-specific responses by binding to CD28 on T cells. The addition of an anti-CD28 mAb to the cell culture enhances antigen-specific responses and can counteract capture effects. For example, the presence of IL-2 capture antibodies may result in reduced activation of T cells, as capturing of IL-2 decreases the amount of available IL-2 and thereby dampens the IL-2-mediated stimulation of T cells. The addition of anti-CD28 mAb restores IFN-γ responses (depicted in the below images). Further optimization may be necessary, depending on the cells and stimuli used. Too high concentrations of the anti-CD28 mAb may result in an elevation of non-specific cytokine secretion.

Overcoming capture effects using anti-CD28 anitbodies in FluoroSpot

(1) An anti-CD28 antibody can be added to provide a co-stimulatory signal that can restore (2) e.g. IFN-γ responses.

How to investigate capture effects

Our FluoroSpot Plus kits are evaluated for capture effects, and in studies of T cell responses we recommend co-stimulation with anti-CD28. With FluoroSpot Flex, it is possible to combine and build your own kit. For guidance look at our analyte combination table (above). Capture effects can be investigated by quantifying spot numbers in wells coated with single capture antibodies and compared to wells coated with a mixture of the capture antibodies. The compensatory effect of the anti-CD28 mAb may be assessed by comparing cells cultured with and without the anti-CD28 mAb.

Species reactivities of our veterinary assays and included monoclonal antibodies are summarized in the tables below.

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

Blank boxes

Represent cross-reactivities not evaluated.

IFN-γ

ELISA and ELISpotCowSheepGoatHorseLlama  Pig  DogFerretRhinocerosSalmonRainbow trout
Bovine IFN-γ
MT17.1/MT307, biotin
( )( )   
Canine IFN-γ
MT13/MT166, biotin
 ( )( )( )( )    
Equine IFN-γ
MT166/MT13, biotin
 ( )( )( )( ) ( )  
Porcine IFN-γ
pIFNγ-I/P2C11, biotin
      
Ferret IFN-γ
MTF14/MTF19, biotin
   
Salmon IFN-γ
MT36X/MT10, biotin
         ( )
Flow Cytometry
mAb MT307        
mAb P2C11          
mAb MTF14          
mAb MT166          

IgG

ELISA and ELISpotCowSheepGoatHorsePigDog
Bovine IgG
ELISpot MT134/MT391, biotin​
ELISA MT134/MT391, HRP​
  
Porcine IgG
ELISpot MT421/MT424, biotin
ELISA MT421/MT424, HRP​
 
Equine IgG
ELISA MT329A4/ MT310A61, biotin​​
  

IL-2

ELISA and ELISpotCowSheepGoatHorsePigDogFerret
Bovine IL-2
MT11A31/MT3B3, biotin
     
Porcine IL-2
MT264/267 /MT265, biotin
     
Ferret IL-2 ELISA
MT264/MT265, biotin
      
Ferret IL-2 ELISpot
MT264/271 /MT265, biotin
      
Flow Cytometry
mAb MT3B3     

IL-4

ELISA and ELISpotCowSheepGoatHorsePigDog
Bovine IL-4
bIL4-I/bIL4-II, biotin
( )

IL-5

ELISA and ELISpotCowSheepGoatHorsePigDog
Bovine IL-5
MT52B34/MT11B37, biotin
   
Porcine IL-5
MTAL25/MTAL59, biotin
     

IL-8

ELISA and ELISpotCowSheepGoatHorsePigDog
Bovine IL-8
MT8H6/26E5, biotin (ELISA)
MT8H6/8F19 /26E5, biotin (ELISpot)
 

IL-10

ELISA and ELISpotCowSheepGoatHorsePigDog
MT3515/MT32AD, biotin   

IL-17A

ELISA and ELISpotCowSheepGoatHorsePigDog
MT49A7/MT51B8, biotin   
MT49A7/MTP853, biotin     
Flow Cytometry
mAb MT504

Perforin

Flow CytometryCowSheepGoatHorsePigDog
mAb Pf-344

TNF-α

ELISA and ELISpotCowSheepGoatHorsePigDog
Bovine TNF-α
MTF4918/MT771, biotin
   
Porcine TNF-α
MT1P14/MT859, biotin (ELISA)
MT32E9/MT859, biotin (ELISpot)
    
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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
Cell typeT cell, Tc, Th1, NK cell
Gene ID281237, 443396