SHOP

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

Human PBMC from a COVID-19 convalescent individual and a healthy control 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, followed by anti-WASP-HRP and the number of cells secreting RBD-specific IgG was analyzed by ELISpot

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

Components

PlatePre-coated ELISpot plate, white MSIP (mAbs MT91/145)
Detection mAbMT78/145, biotin
AntigenRBD-WASP
Enzyme conjugatesStreptavidin-HRP enzyme conjugate
Anti-WASP mAb, HRP
SubstrateTMB substrate for ELISpot
Buffer/SolutionReconstitution buffer

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 (HRP)
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 and a healthy control 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, followed by anti-WASP-HRP and the number of cells secreting RBD-specific IgG was analyzed by ELISpot
Human PBMC from a COVID-19 convalescent individual and a healthy control 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-HRP and the number of cells secreting RBD-specific IgG was analyzed by ELISpot.

Tutorial

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

B cell profiling with ELISpot and FluoroSpot

ELISpot and FluoroSpot are considered the gold standard method for finding rare, antigen-specific B cells.
Tutorial

Step-by-step guide to ELISpot

An in-depth guide to ELISpot, including best practice and tips and tricks.
Tutorial

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

What is a positive ELISpot response?

DFR is a non-parametric statistical test for determining positive responses in ELISpot and FluoroSpot.
Tutorial

Isolation, freezing, and thawing of PBMCs

This is a protocol on how to best isolate, freeze, and thaw PBMCs.

Highlighted research

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

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

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.

1 / 1

ELISpot Path: SARS-CoV-2 (S1scan+SNMO) Human IFN-γ (HRP)