Anti-human IL-12 (p70) mAb (MT704), biotin

Anti‑human IL‑12 (p70) mAb (MT704), biotin

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Content

Monoclonal antibody MT704, biotinylated. Supplied at 0.5 mg/ml in PBS with 0.02% sodium azide.

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Product specifications

Intended use

This monoclonal antibody enables specific detection of human IL-12 (p70) in immunoassays such as ELISpot, FluoroSpot, and ELISA.

Serum/Plasma samples

Recommendation

MT704 is recommended as detection mAb in ELISpot, FluoroSpot, and ELISA in combination with capture mAbs MT86/221 (product code 3450-3).

Product details

ProductAnti-human IL-12 (p70) mAb (MT704), biotin
ApplicationELISpot, FluoroSpot, ELISA
AnalyteIL-12 (p70)
AntibodyMT704
ConjugateBiotin
ClonalityMonoclonal
ImmunogenRecombinant human IL-12 p70
HostMouse
IsotypeIgG2b
ReactivityHuman, Non-human primates
Specificity

Native and recombinant human IL-12 p70

Cross-reactivity

Cross-reacts with IL-12 p70 from cynomolgus macaque. For information on reactivity with other non-human primates (NHP), please check the NHP cross-reactivity guide.

Purification

Purified from in vitro cultures by protein G affinity chromatography.

Biotinylation

Biotinylated through reaction with a N-hydroxysuccinimide ester of biotin.

Concentration0.5 mg/ml
Supplied in

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

Contents

Monoclonal antibody MT704, biotinylated. 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.
Datasheet/Protocol
SDS

Learn how to combine Mabtech’s antibodies for the detection of human IL-12, IL-12/-23 (p40), IL-23, IL-27, and EBI3 in ELISA and ELISpot.

 

The IL-12 cytokine family

The IL-12 cytokine family consists of IL-12, IL-23, IL-27, IL-35, and IL-39. The existence of IL-39 in human remains, however, to be confirmed. The proteins are heterodimers of different subunits which are either unique or shared by several family members. IL-12 is a heterodimer, (p70), consisting of two covalently linked subunits, p35 and p40. The p40 subunit is shared with IL-23 which is a heterodimer of p40 and a unique p19 subunit. The p40 subunit also exists as a monomer and homodimer. The EBI3 subunit is shared with IL-27, IL-35, and IL-39.

Mabtech supplies monoclonal antibodies with defined subunit specificities to p40, p35, p19, p28, and EBI3. Since both IL-12 and IL-23 heterodimers contain the p40 subunit, capture immunoassays based on antibodies to p40 will detect both cytokines. Such assays can also detect p40 monomers and dimers. The EBI3 assays will detect IL-27, IL-35, and IL-39.

In our IL-12 and IL-23 assays, the same capture antibodies to p40 (MT86/221) are used. Different detection antibodies enable the detection of IL-12 (p70), IL-12/-23 (p40), and IL-23.

 

Application guide: mAbs to human IL-12, IL-23, IL-27 and EBI3

 

 IL-12 (p70) 
 
 
Capture mAbs: MT86/221 (p40 specific)
 
Detection mAb: MT704 (p70 specific)

     

 IL-23 
 
 
Capture mAbs: MT86/221 (p40 specific)
 
Detection mAb: MT155 (p19 specific)

     

 IL-27 
 
 
Capture mAbs: MT27/361 (IL-27 and p28 specific)
 
Detection mAb: MT140 (EBI3 specific)
 IL-12/-23 (p40) 
 
 
Capture mAbs: MT86/221 (p40 specific)
 
Detection mAb: MT618 (p40 specific)

     

 EBI3 (IL-27 and IL-35) 
 
Capture mAbs: MT23A14 (EBI3 specific)
 
Detection mAb: MT140 (EBI3 specific)

 

 

Get inspired!

Make sure to check out how researchers have used Mabtech's IL-23 ELISA, ELISpot, and FluoroSpot kits in everything from infectious disease research to novel immunotherapies. You can find it all in our Publication database.

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

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.

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)( )  ( )( )( )( )
TNF-α       
          
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 December 28, 2023

Incubation strategies for success in ELISpot and FluoroSpot
We've made a handy table summarizing incubation times for different analytes to make your research planning for the ELISpot and FluoroSpot assay a little easier.
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IL-12 (p70)

Analyte description

Interleukin 12 (IL-12) is, when biologically active, a heterodimer (p70) consisting of two covalently linked subunits, p35 and p40. The p40 subunit is shared with IL-23. IL-12 is produced by antigen-presenting cells, specifically dendritic cells, and macrophages. IL-12 is a proinflammatory cytokine that promotes Th1-type responses by inducing IFN-γ production and enhancing the proliferation and cytotoxicity of NK and T cells.

Alternative namesInterleukin 12, IL-12, IL12, CLMF, IL-12A, NFSK, NKSF1, P35, CLMF2, IL-12B, IMD28, IMD29, NKSF, NKSF2
Cell typeMonocyte/MΦ, mDC
Gene ID3592, 3593
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ELISpot Plus: Human IL-12 p70 (ALP)