Let Creative Biolabs Help You with FluoroSpot Assay

The FluoroSpot assay at Creative Biolabs has been developed as an alteration of colorimetric substrates-based traditional enzymatic ELISpot detection systems. ELISpot is a good choice for the functional analysis of single immune cells, while FluoroSpot can be used for the functional analysis of 7 subsets of immune cells. The FluoroSpot is capable of detecting secreted cytokines or antibodies with a high level of sensitivity with the use of fluorochrome-conjugated detection antibodies.

Fig.1 6 General steps of FluoroSpot.

The most common cytokines investigated are IFN-ɣ, IL-2, IL-5, IL-10, IL-17, TNF, GM-CSF, Granzyme B, etc.

Fig.2 Most common cytokines investigated.

We also provide B cell FluoroSpot assays for:

• The analysis of human IgG in response to spike protein of SARS-CoV-2
• The analysis of human IgG in response to RBD of SARS-CoV-2
• The analysis of human IgG in response to spike protein and RBD of SARS-CoV-2
• The analysis of mouse IgA and IgG
• The analysis of mouse IgG and IgM
• The analysis of mouse IgA, IgG, and IgM

Advantages

• The highly effective tool with a high sensitivity
• Detection of two or more different cytokines at the sample time
• The lower amount of PBMCs needed
• It can be used to evaluate B-cell and T-cell responses in individuals.
• It can be potentially used for clinical trial monitoring.

Publications Sharing Based on FluoroSpot Assays

Publication 1

Cells Used: Peripheral blood mononuclear cells (PBMCs)

Journal: Methods in molecular biology

IF: 0.368

Research Findings: PBMCs were stimulated with phorbol myristate acetate (PMA) and ionomycin in PVDF plates, and then, IFN-γ- and/or IL-2-producing cells were analyzed by a dual-color fluorospot assay. Results suggest the broad use of FluoroSpot assay for characterization of subgroups of T cells and polarization antigen-specific T cells.

Fig.3 Dual-color fluorospot for the detection of IL-2 and/or IFN-γ-producing cells.¹

Publication 2

Cells Used: Peripheral blood mononuclear cells (PBMCs)

Journal: Journal for ImmunoTherapy of Cancer

IF: 10.9

Research Findings: PBMCs were analyzed for the secretion of IFN-ɣ, IL-17A, and IL-22 by FluoroSpot. 7 sub-populations, including 3 single secretors, 3 dual secretors, and 1 triple secretor, were identified by multi-level evaluation.

Fig.4 Overlay image of a triple color FluoroSpot well.²

Scientists at Creative Biolabs have first-class technology and offer comprehensive service in applying FluoroSpot assay.

For more detailed information about our FluoroSpot Service, please contact us immediately.

References

1. Gazagne, Agnes, et al. "Fluorospot assay: methodological analysis." Handbook of ELISPOT: Methods and Protocols (2005): 289-295.
2. Janetzki, Sylvia. "Immune monitoring technology primer: the enzyme-linked immunospot (Elispot) and Fluorospot assay." Journal for Immunotherapy of Cancer 3.1 (2015): 30.

For Research Use Only | Not For Clinical Use