IFNg Mouse Antibody for Flow Cytometry

Biological Sciences Platform
Sunnybrook Research Institute
Postdoctoral Researcher

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Company:

BioLegend

Product Name:

PE anti-mouse IFN-γ Antibody

Catalog Number:

505808

Clone Number:

XMG1.2

I am researching immunotherapies for cancer in mice, and my general research aim is to determine the extent of immune cell infiltration as well as activation by staining cytokines such as IFNg. For this experiment, C57Bl/6 splenocytes were stimulated for 6 hours with PMA/ionomycin vs splenocytes that were not stimulated. Cells were stained with CD8 PE-Cy7 (Biolegend 100722) to label cytotoxic T-cells and IFNg to label activated CD8+ T-cells. As a control, unstimulated cells and cells that had been stained with all antibodies except IFNg-PE were used (Fluorescence minus one (FMO)).

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

C57Bl6 splenocytes

Primary Incubation

1:100 dilution on ice for 1 hour

Blocking Agent

BSA

Secondary Incubation

None

Tertiary Incubation

None

Detection

Flow cytometry

Results Summary

Upon PMA/ionomycin stimulation the percentage of CD8+ T-cells expressing IFNg increased from 0.4% to 5.65%. FMO controls are included of IFNg-PE to show gating strategy.

DOI or PMID #

N/A

Additional Notes

Cells were fixed and permeabilised using eBioscience FoxP3 staining kit.

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Summary

The Good

Robust bright antibody of a well characterised clone

The Bad

None

The Bottom Line

This is a useful antibody to include in panels where IFNg expression is being investigated.

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