Great Flow Antibody to Detect IFNg Secretion in T Cells

Oncology
Johns Hopkins School of Medicine
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Fisher Scientific

Product Name:

IFN gamma Monoclonal Antibody (XMG1.2), PerCP-Cyanine5.5, eBioscience

Catalog Number:

45-7311-82

As an immunologist, we assess CD8 T cell cytokine secretion via flow cytometry. We analyze interferon gamma secretion (IFNg) since this cytokine is one of the first cytokines to be expressed for cytolytic activity. Upon stimulation, we can detect expression of this cytokine in CD8 T cells. The antibody is great to detect IFNg expression in T cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine T-cells during an infection

Primary Incubation

1:500 in 1x BD perm/wash buffer after surface staining and fix/perming

Blocking Agent

FC block 100 µg/mL in FACS buffer

Secondary Incubation

None

Tertiary Incubation

None

Detection

BD Celesta flow analyzer

Results Summary

We are able to detect IFNg expression in T cells by re-stimulating T cells in the presence of a golgi inhibitor to prevent secretion.

DOI or PMID #

N/A

Additional Notes

Make sure to surface stain then fix/perm (BD Bioscience) then stain ICS in 1x Perm Wash

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Summary

The Good

Really good and bright antibody to detect IFNg expression along with other cytokines.

The Bad

None whatsoever

The Bottom Line

Great antibody and clone to detect IFNg expression in T cells upon stimulation.

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