ELISpot Flex: Mouse IgM (ALP)

ELISpot Flex: Mouse IgM (ALP)

3885-2A

Publications: 17
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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 mAbMT6A3
Detection mAbMT9A2, biotin
Enzyme conjugateStreptavidin-ALP
StimuliR848
Recombinant mouse IL-2

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Delivery 4-9 business days

Shipping $0

Complementary products

Complementary products
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ELISpot plates, white


Product specifications

Intended use

This ELISpot Flex kit is intended for the enumeration of B cells secreting mouse 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 mouse IL-2 (both included).

Recommendation

Product details

ProductELISpot Flex: Mouse IgM (ALP)
ApplicationELISpot
AnalyteIgM
ReactivityMouse
Specificity

This kit is based on a matched pair of mAbs enabling specific detection of mouse IgM.

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.

Mouse spleen cells (naïve cells) were pre-incubated for 72 h in the absence or presence of R848 and recombinant IL-2. The cells were then washed, added into ELISpot wells and analyzed for the secretion of total IgM (30,000 cells/well, overnight incubation).

Datasheet/Protocol
SDS

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.

Tutorial, Published November 24, 2022

Cell stimulation in ELISpot and FluoroSpot
In this feature, we’ll walk you through the optimal use of different polyclonal and antigen-specific stimuli.

Tutorial, Published November 21, 2022

B cell profiling with ELISpot and FluoroSpot
ELISpot and FluoroSpot are considered the gold standard method for finding rare, antigen-specific B cells.

Tutorial, Published November 7, 2022

7 common ELISpot errors and how to avoid them
ELISpot troubleshooting. We discuss 7 of the most common issues we’ve encountered in ELISpot, what mistakes causes them and how to avoid and correct them.

Tutorial, Published October 14, 2022

What is a positive ELISpot response?
DFR is a non-parametric statistical test for determining positive responses in ELISpot and FluoroSpot.

Tutorial, Published October 14, 2022

Isolation, freezing, and thawing of splenocytes
This is a protocol on how to best (i) isolate, (ii) freeze, and (iii) thaw splenocytes.
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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, AI326478, Igh-6, Igh-M, Igh6, Igm, muH
Cell typeB cell
Gene ID16019