Intracellular Staining

August 13, 2015

Immunology
University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

eBiosciences

Product Name:

Anti-Mouse CD279 (PD-1) APC

Catalog Number:

17-9981

Info: View Product View Product Specs

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We investigated the expression and upregulation of coinhibitory molecules on antigen-specific T cells as they undergo deletional tolerance.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Murine T cells from lymph nodes and spleen

Primary Incubation

1:100 for 30 minutes at 4 degC after cytofix/cytoperm

Blocking Agent

2.4G2 Fc block

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Canto II flow cytometer

Results Summary

We used the BD Biosciences cytofix/cytoperm kit for intracellular staining. The PD-1 APC antibody gave strong staining, and we could clearly see that PD-1 was upregulated as the cells proliferated, as expected. We tested other PD-1 clones and colors and found the best results using either PE or APC of the RMP1-14 clone.

Additional Notes

None

Image Gallery

Summary

The Good

Good, clear staining

The Bad

Requires high antibody concentration and intracellular staining

The Bottom Line

One of the best antibodies available for PD-1 staining; strong staining when used at 1:100 intracellularly. Best with an isotype control to identify any background staining.

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