Detection of Mouse Intestinal Macrophages by IFA

October 28, 2015

Microbiology and Immunology
Loyola University Chicago
Research Associate

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Company:

BioLegend

Product Name:

PE-anti-mouse F4/80

Catalog Number:

123109

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To analyze the activation of mouse intestinal macrophages after experiment treatment, mice were gavaged with reagent and intestinal tissue was collected after 24 hours. Intestinal tissue was snap frozen and cryo sections were used for staining the intestinal macrophages. We found intestinal macrophages (F4/80 +) and also different activation status of the macrophages. This anti-mouse F4/80 antibody is enough to generate signal for visualizing the macrophage in LP or peyer's patches.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Intestinal tissue

Primary Incubation

PE-anti-mouse F4/80 with 1:50 to 100 dilution

Blocking Agent

5% FBS in PBS

Secondary Incubation

NA

Tertiary Incubation

NA

Detection

Slides were coverslipped with DYPI containing mounting medium and F4/80 + cells were analyzed under flourescence microscropy

Results Summary

Results showed that F4/80 positive cells could be easily found in the intestine LP and peyer’s patch (figure shows macrophages in mouse peyer's patch).

Additional Notes

To generate good signal, slides need to be blocked at least 1 hour and antibody needs to be tested at different dilutions.

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Summary

The Good

One step staining is able to get adequate signal to see F4/80 +cells

The Bad

Signal is not as good as the antibody with 2 step staining

The Bottom Line

This antibody is able to be used for IFA and can obtain enough signal for visualizing the macrophage in LP or peyer's patch. We have compared it with primary anti-F4/80antibody that needs labeled 2nd antibody, the latter generates stronger signal. However, one step staining can save you time.

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