ELISpot Flex: Human IgA (ALP)

ELISpot Flex: Human IgA (ALP)

3860-2A

Publications: 16
Documents
Questions?
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 mAbMT57
Detection mAbsMT20, biotin
MT57, biotin
Enzyme conjugateStreptavidin-ALP
StimuliR848
Recombinant human IL-2

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Delivery 4-9 business days

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ELISpot plates, white


Product specifications

Intended use

This ELISpot Flex kit is intended for the enumeration of B cells secreting human IgA using the ELISpot assay. B cells secreting antigen-specific IgA, as well as B cells secreting IgA irrespective of antigen specificity (total IgA) 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).

Recommendation

Product details

ProductELISpot Flex: Human IgA (ALP)
ApplicationELISpot
AnalyteIgA
ReactivityHuman
Specificity

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

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 into ELISpot wells and analyzed for the secretion of total IgA (50,000 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 September 2, 2022

Isolation, freezing, and thawing of PBMCs
This is a protocol on how to best isolate, freeze, and thaw PBMCs.

Highlighted research, Published May 30, 2020

B cell FluoroSpot for sensitive ASC detection
Mabtech IRIS and IgG/IgA/IgM FluoroSpot were used to study antibody-secreting plasma cells (ASCs) specific for the nucleocapsid (N) protein. FluoroSpot was shown to be more sensitive than e.g., ELISA.

Highlighted research, Published October 15, 2019

Detection of tumor infiltrating B cells
Tumor infiltrating B cells are able to actively secrete immunoglobulins as shown by performing an IgA, IgG and IgM FluoroSpot. The FluoroSpot technology enabled investigation of the functionality of cells directly isolated from the tumor.
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IgA

Analyte description

IgA (Immunoglobulin A) represents only 10-15% of the total Ig molecules in serum but is the predominating Ig isotype in secretions and in the gastrointestinal and urogenital tract. The IgA found in secretions (secretory IgA or sIgA) is commonly a dimer with two IgA molecules joined by the J chain. Secretory IgA is the most important Ig isotype in mucous surfaces, the area of entry for most pathogens.

Alternative namesImmunoglobulin A, IgA, A2M, IgA1
Cell typeB cell
Gene ID3494, 3493