Bright Stain for Interferon Gamma

Pathology
Brigham and Women's Hospital
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

Alexa fluor 647 anti-mouse IFN-gamma

Catalog Number:

557735

This antibody binds to mouse interferon gamma, a cytokine that is involved in T cell immunity and inflammation. Since, cytokines have a short half-life and can be consumed during signaling, they are hard to detect. By blocking ER transport for a few hours along with strong re-stimulation, some antibodies can detect IFNg very well. However, this antibody gave a bright fluorescence which was well- separated from the negative population.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Purified mouse T cells

Primary Incubation

1:200 in Perm/Wash buffer

Blocking Agent

FcR block

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow Cytometry - APC channel

Results Summary

The cells show bright IFNg staining which i haven't observed with other fluorophore conjugated antibodies. Background on unstimulated cells is minimal.

Additional Notes

None

Related Categories

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Summary

The Good

Bright stain and easy to use

The Bad

Needs restimulation followed by intracellular staining

The Bottom Line

It requires a long protocol for staining, but gives good results

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