ELISpot Path: SARS-CoV-2 (RBD) Human IgG (ALP)

ELISpot Path: SARS‑CoV‑2 (RBD) Human IgG (ALP)

3850-4APW-R1-1

Publications: 22
Documents
Questions?
A straightforward setup for functional analysis of a specific scientific question.

Components

PlatePre-coated ELISpot plate, white MSIP (mAbs MT91/145)
Detection mAbMT78/145, biotin
AntigenRBD-WASP
Enzyme conjugatesStreptavidin-ALP
Anti-WASP mAb, ALP
SubstrateBCIP/NBT-plus
Buffer/SolutionReconstitution buffer

In stock

Delivery 4-9 business days

Shipping $0

Product specifications

Intended use

This ELISpot Path kit is intended for the enumeration of B cells secreting human IgG in response to SARS-CoV-2 (RBD) using the ELISpot assay. B cells secreting IgG irrespective of antigen specificity (total IgG) can also be enumerated. For research use only. Not for use in diagnostic procedures.

Serum/Plasma samples

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

Recommendation

Product details

ProductELISpot Path: SARS-CoV-2 (RBD) Human IgG (ALP)
ApplicationELISpot
AnalyteIgG
ReactivityHuman
Specificity

This kit is based on a matched pair of mAbs specific for the Fc part of human IgG.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store antibodies, enzyme conjugate, and substrate at 4-8 °C. Recombinant protein should be stored at -20 °C or below. Plates may be kept at room temperature.

Shelf lifeAt least 12 months from date of receipt.
Human PBMC from a COVID-19 convalescent individual were incubated in the presence and absence of R848+IL-2 (stimulates memory B cells) in tubes for three days. Cells were then extensively washed and added to an ELISpot plate (200,000 cells/well) coated with anti-human IgG mAbs. RBD-specific spots were detected with RBD-WASP, i.e., recombinant Receptor Binding Domain protein with a WASP-tag, followed by anti-WASP-ALP and the number of cells secreting RBD-specific IgG was analyzed by ELISpot.
Datasheet/Protocol
SDS

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 December 16, 2021

SARS-CoV-2-specific immune responses in patients with autoimmune diseases
Patients with rheumatic diseases showed T cell responses comparable to healthy control. Patients with MS and after mRNA vaccination show different memory B cell IgG responses. SARS-CoV-2 specific immune responses in autoimmunity was analyzed using FluoroSpot, ELISpot Path, and Mabtech IRIS.

Highlighted research, Published December 2, 2021

Importance of studying SARS-CoV-2-specific memory B cells
When studying antibodies in circulation, remember that memory B cells might hide out of sight. In this study, detection of SARS-CoV-2-specific T and memory B cells 6 and 15 months after infection.

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.
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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, IgG3
Cell typeB cell
Gene ID3500, 3501, 3502