Paves your way to functional analysis of a specific research question.
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.
For memory B cells, we recommend pre-stimulation with R848 and recombinant human IL-2 (both included).
Performance
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.
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.