ELISpot Flex: Human IgG4 (ALP)

3854-2A

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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 mAb	MT145
Detection mAb	MTG42, biotin
Enzyme conjugate	Streptavidin-ALP
Stimuli	R848
	Recombinant human IL-2

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Delivery 4-9 business days

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Anti-human IgG4 mAb (MTG42), unconjugated

Product specifications

Intended use

This ELISpot Flex kit is intended for the enumeration of cells secreting human IgG4 using the ELISpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

With this kit, detection of antigen-specific IgG4 secreting B cells can be performed using coated antigen. If detection with biotinylated antigen is preferred, the plate should instead be coated with the subclass specific antibody.

For memory B cells, we recommend pre-stimulation with R848 and recombinant human IL-2 (both included).

Recommendation

Product details

Product	ELISpot Flex: Human IgG4 (ALP)
Application	ELISpot
Analyte	IgG4
Reactivity	Human
Specificity	This kit is based on a matched pair of mAbs enabling specific detection of human IgG4.

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 life	At least 18 months from date of receipt.

Performance

PBMC were pre-incubated for 4 days in the presence or absence of R848 and recombinant IL-2. The cells were then washed, added into ELISpot wells (stimulated cells: 200,000 cells/well, unstimulated cells: 100,000 cells/well) and analyzed for the secretion of total IgG4 (overnight incubation).

Documents

Datasheet/Protocol
SDS

Human IgG4 mAb guide

Selecting the appropriate mAbs is key to detecting both total and antigen-specific human IgG4. This blog post explores Mabtech’s mAbs, tailored for ELISpot to detect human IgG4 antibody-secreting cells.

Application guide: ELISpot

Ag-specific IgG4

Coating:
Antigen
Detection mAb:
MTG42, biotin

Coating:
MTG42, unconjugated
Detection mAb:
Antigen, biotin

Total IgG4

Coating:
MT145, unconjugated
Detection mAb:
MTG42, biotin

Tutorials

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.

Publications (0)

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Analyte information

IgG4

Analyte description	IgG (Immunoglobulin G) is the most abundant Ig isotype in serum, making up approximately 80% of all serum Immunoglobulins. In humans, there are four subclasses of IgG, with the highest serum concentrations of IgG1 followed by IgG2, IgG3, and IgG4. IgG4 cannot activate the complement system but binds well to Fc receptors.
Alternative names	Immunoglobulin G4, IgG4
Cell type	B cell
Gene ID	3503