ELISpot Flex: Human IgM (ALP)

ELISpot Flex: Human IgM (ALP)

3880-2A

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Our most adaptable ELISpot format, made to fit into your unique plan. Combine it with a substrate, plate, and/or antigen of choice.

Components

Capture mAbMT11/12
Detection mAbMT22, biotin
Enzyme conjugateStreptavidin-ALP
StimuliR848
Recombinant human IL-2

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Product specifications

Intended use

This ELISpot Flex kit is intended for the enumeration of B cells secreting human IgM using the ELISpot assay. B cells secreting antigen-specific IgM, as well as B cells secreting IgM irrespective of antigen specificity (total IgM) can be enumerated. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

Antigen-specific B cells can be enumerated in two alternative ways; by coating with antigen or by using biotinylated antigen in the detection step. For memory B cells, we recommend pre-stimulation with R848 and recombinant human IL-2 (both included).

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Product details

ProductELISpot Flex: Human IgM (ALP)
ApplicationELISpot
AnalyteIgM
ReactivityHuman, Non-human primates
Specificity

This kit is based on a matched pair of mAbs specific for human IgM.

Cross-reactivity

The mAbs react with human IgM and cross-react with IgM from non-human primates (NHP). For more information please check the Cross-reactivity guide section below.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store antibodies and enzyme conjugate at 4-8 °C upon receipt. R848 and IL-2 should be stored at -20 °C.

Shelf lifeAt least 18 months from date of receipt.
PBMC were pre-incubated for 72 h in the absence or presence of R848 and recombinant IL-2. The cells were then washed, added to an ELISpot plate and analyzed for the secretion of total IgM (50,000 cells/well, overnight incubation).

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 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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IgM

Analyte description

Immunoglobulin M (IgM) is the first antibody isotype (also called class) secreted by the adaptive immune system in response to a foreign antigen. Plasma B cells secrete IgM mainly as pentamers, i.e., five IgM molecules joined together by the J chain. Because IgM is formed early in the immune response and is later replaced by IgG, specific antibodies of the IgM class are characteristic of a recent, ongoing, or chronic infection.  Therefore, IgM represents the isotype with the lowest affinity for antigens. Due to its structure, with 10 antigen-binding sites per IgM pentamer, it can nevertheless bind surface-bound antigens with very high avidity. Immunoglobulin M constitutes approximately 10% of the total immunoglobulin concentration in serum. Resting mature yet naive, B lymphocytes express monomeric IgM as a transmembrane antigen receptor that functions as part of the B cell receptor (BCR).

Alternative namesImmunoglobulin M, IgM, AGM1, MU, VH
Cell typeB cell
Gene ID3507