SHOP

FluoroSpot Path: SARS‑CoV‑2 (Spike​+​RBD) Human IgG

Human PBMC from a COVID-19 convalescent individual were incubated with R848+IL-2 (stimulates memory B cells) in tubes for three days. Cells were then extensively washed and added to an FluoroSpot plate (250,000 cells/well) coated with anti-human IgG mAbs. Spike-specific IgG spots were detected with Spike-GAL, i.e., recombinant Spike protein with a GAL peptide tag, followed by anti-GAL-490. RBD-specific spots were detected with RBD-WASP, i.e., recombinant Receptor Binding Domain protein with a WASP peptide t

FluoroSpot Path: SARS‑CoV‑2 (Spike​+​RBD) Human IgG

Components

PlatePre-coated FluoroSpot plate (mAbs MT91/145)
Detection mAbAnti-IgG mAbs (MT78/145), 550
AntigensSpike-GAL
RBD-WASP
Fluorophore conjugatesAnti-WASP mAb, 640
Anti-GAL mAb, 490
Buffers/SolutionsFluoroSpot enhancer
Reconstitution buffer
StimuliR848
Recombinant human IL-2

Intended use

This FluoroSpot Path kit is intended for the enumeration of B cells secreting human IgG in response to SARS-CoV-2 (Spike and RBD) using the FluoroSpot 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

ProductFluoroSpot Path: SARS-CoV-2 (Spike+RBD) Human IgG
ApplicationFluoroSpot
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, fluorophore conjugates, and FluoroSpot enhancer at 4-8 °C upon receipt. Recombinant proteins 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 with R848+IL-2 (stimulates memory B cells) in tubes for three days. Cells were then extensively washed and added to an FluoroSpot plate (250,000 cells/well) coated with anti-human IgG mAbs. Spike-specific IgG spots were detected with Spike-GAL, i.e., recombinant Spike protein with a GAL peptide tag, followed by anti-GAL-490. RBD-specific spots were detected with RBD-WASP, i.e., recombinant Receptor Binding Domain protein with a WASP peptide t
Human PBMC from a COVID-19 convalescent individual were incubated with R848+IL-2 (stimulates memory B cells) in tubes for three days. Cells were then extensively washed and added to an FluoroSpot plate (250,000 cells/well) coated with anti-human IgG mAbs. Spike-specific IgG spots were detected with Spike-GAL, i.e., recombinant Spike protein with a GAL peptide tag, followed by anti-GAL-490. RBD-specific spots were detected with RBD-WASP, i.e., recombinant Receptor Binding Domain protein with a WASP peptide tag, followed by anti-WASP-640. Spots were analyzed with Mabtech IRIS.

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 FluoroSpot

An in-depth guide to FluoroSpot, including best practice and tricks we've learnt over the years.
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.