FluoroSpot Flex: Mouse IgM/IgG

FluoroSpot Flex: Mouse IgM/IgG

FS-60G49R-1

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This FluoroSpot kit can be set up in many ways – designed for B cell profiling.

Components

PlateFluoroSpot plate (IPFL), non-coated
Capture mAbMT24/JC5-1
Detection mAbsAnti-IgM mAb (MT9A2), 490
Anti-IgG mAbs (MG1/MG2a/MTG2c/MG2b/MG3), 550
Buffer/SolutionFluoroSpot enhancer
StimuliR848
Recombinant mouse IL-2

In stock

Delivery 4-9 business days

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Product specifications

Intended use

This FluoroSpot Flex kit is intended for the enumeration of B cells secreting mouse IgM and mouse IgG using the FluoroSpot assay. B cells secreting antigen-specific antibodies as well as B cells secreting antibodies irrespective of antigen specificity can be measured with this kit. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

For analysis of memory B cells, cells may need to be pre-activated with R848 and recombinant mouse IL-2 (both included).

Recommendation

Product details

ProductFluoroSpot Flex: Mouse IgM/IgG
ApplicationFluoroSpot
AnalyteIgM, IgG
ReactivityMouse
Specificity

This kit is based on mAbs enabling specific detection of mouse IgM and mouse IgG.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store antibodies, fluorophore conjugates, and FluoroSpot enhancer at 4-8 °C upon receipt. R848 and IL-2 should be stored at -20 °C. Plates may be kept at room temperature.

Shelf lifeAt least 12 months from date of receipt.

Mouse IgM and IgG images from analysis of in vivo activated splenocytes from hyperimmunized (with ovalbumin) BALB/c mice in IgM/IgG/IgA FluoroSpot. 75,000 cell/well were incubated in the FluoroSpot plate for 5 hours without stimulation.

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.
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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
Cell typeB cell

IgG

Analyte description

Immunoglobulin G (IgG) is the most abundant Ig isotype in serum, making up approximately 80% of all serum immunoglobulins. In humans, there are four subclasses of immunoglobulin G, with the highest serum concentrations of IgG1 followed by IgG2, IgG3, and IgG4. In mice, the IgG subclasses are defined as IgG1, IgG2a/c, IgG2b, and IgG3. The IgG molecule consists of two heavy and two light chains (κ or λ), resulting in a molecule with two arms for antigen binding. High levels of IgG antibodies are induced following the initial IgM response in a typical immune response to antigens.

Alternative namesImmunoglobulin G, IgG
Cell typeB cell