Great Antibody to Detect TNF Alpha Expression

Pharmacology and Oncology
Johns Hopkins School of Medicine
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

BD Biosciences

Product Name:

PE Rat Anti-Mouse TNF

Catalog Number:

554419

As an immunology lab studying T cells, we routinely assess cytokine expression of canonical cytokines such as TNF alpha. Using flow cytometry, we stimulate T cells and interrogate expression of cytokines such as TNF alpha. This is an important cytokine since it plays a very critical role in the effector response.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

In vitro murine CD8 T cells

Primary Incubation

1:500 in 1x BD perm wash (intracellular staining)

Blocking Agent

Surface staining FC blocking (10 µg/mL)

Secondary Incubation

None

Tertiary Incubation

None

Detection

BD Celesta

Results Summary

We are readily able to determine TNF alpha expression from cytokines based on proliferation. Cells that express TNF alpha are easily detected as positive whereas non producers have minimal expression.

DOI or PMID #

N/A

Additional Notes

Remember to surface stain with right markers and then proceed with BD Cytofix/Perm kit to fix and perm cells before intracellular staining.

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Summary

The Good

Nice cheap antibody since its PE to stain for TNF alpha expression.

The Bad

None really.

The Bottom Line

Reliable and robust antibody to detect TNF alpha in lymphocytes upon stimulation for flow cytometry.

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