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

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

FSP-05R-RS1-1

Publications: 0
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Paves your way to functional analysis of a specific research question.

Components

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

In stock

Delivery 4-9 business days

Shipping $0

Product specifications

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 tag, followed by anti-WASP-640. Spots were analyzed with Mabtech IRIS.
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 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