Intracellular detection of IFN-gamma

Cancer Immunology & AIDS
Dana Farber Cancer Institute
Research Fellow

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

BioLegend

Product Name:

PerCP/Cy5.5 anti-mouse IFN-γ Antibody

Catalog Number:

505822

Mouse transgenic OT-1 cells were stimulated with their cognate peptide, OVA, for 3 days. On the last day, cells were re-stimulated with PMA/ionomycin and treated with Brefeldin A to allow accumulation of protein in the cytoplasm. Following this, the cells were fixed in 1% formalin and then stained with this anti-mouse IFN-gamma antibody. Intracellular staining of cytokines is a difficult process and takes a lot of standardizing. However, with this antibody the results were obtained easily.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Purified mouse CD8 T cells

Primary Incubation

1:200 dilution for 1 hour on ice

Blocking Agent

NA

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Flow cytometry on PERCP channel

Results Summary

The OVA peptide stimulated cells show increase in IFN-gamma expression, which is expected. The fluorescence shift is not so great, but that maybe attributed to lower expression levels.

Additional Notes

None

Related Categories

Image Gallery

Summary

The Good

It works

The Bad

Lots of standardization is required

The Bottom Line

A great antibody to start learning intracellular staining of cytokines.

Share your experience with other scientists. Write a Review! »

Join the discussion