Adequate Siglec-F staining

February 04, 2016

BlueskyReddit
MBI
University of Rochester
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Miltenyi Biotec

Product Name:

Anti-Siglec-F-PE, mouse

Catalog Number:

130-102-274

Mice were IP injected with zymosan and peritoneal leukocytes were recovered by lavage 2 days later. Total recovered cells were stained to identify eosinophils.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Total Inflammed Peritoneal Leukocytes

Primary Incubation

1:20 anti-SiglecF-PE - 30 mins on ice

Blocking Agent

1:100 FcBlock - 10 mins on ice

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

LSR II (120mW Green Laser)

Results Summary

Eosinophils were clearly identifiable over background.

Additional Notes

Staining at 1:10 and 1:20 gave very similar staining index results.

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Summary

The Good

Reagent comes pre-titrated so optimization is easy. Okay staining intensity.

The Bad

Separation between positive and negative is less than seen with BD siglecF clone on PE

The Bottom Line

Not bad in a pinch but I'll reorder from BD when this vial is done.

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