ELISpot Flex: Mouse IgG3 (ALP)

3826-2A

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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	MT24/JC5-1
Detection mAb	MG3, biotin
Enzyme conjugate	Streptavidin-ALP
Stimulus	R848

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Delivery 4-9 business days

Shipping $0

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Anti-mouse IgG3 mAb (MG3), unconjugated

Product specifications

Intended use

This ELISpot Flex kit is intended for the enumeration of cells secreting mouse IgG3 using the ELISpot assay. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

With this kit, detection of antigen-specific IgG3 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 mouse IL-2 (both included).

Recommendation

Product details

Product	ELISpot Flex: Mouse IgG3 (ALP)
Application	ELISpot
Analyte	IgG3
Reactivity	Mouse
Specificity	This kit is based on a matched pair of mAbs enabling specific detection of mouse IgG3.

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

Splenocytes from C57/BL6 mice 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 IgG3 (50,000 cells/well, overnight incubation).

Documents

Datasheet/Protocol
SDS

Mouse IgG3 mAb guide

Selecting the appropriate mAbs is key to detecting both total and antigen-specific mouse IgG3. This blog post explores Mabtech’s mAbs, tailored for ELISpot to detect mouse IgG3 antibody-secreting cells.

Application guide: ELISpot

Antigen-specific IgG3

Coating:
Antigen
Detection mAb:
MG3, biotin

Coating mAbs:
MT24/JC5-1, unconjugated
Detecting:
Antigen-biotin

Total IgG3

Coating mAbs:
MT24/JC5-1, unconjugated
Detecting:
MG3, 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 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.

Publications (0)

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Analyte information

IgG3

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 IgG, 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. IgG3 binds to Fc receptors with high affinity and is a good complement activator.
Alternative names	Immunoglobulin G3, IgG3
Cell type	B cell
Gene ID	380795