Fluorescent Staining of Mouse Intestinal Epithelial Cells

Microbiology and Immunology
Loyola University Chicago
Research Associate

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Company:

BioLegend

Product Name:

Alexa Fluor® 594 anti-rat IgG

Catalog Number:

405422

To detect mouse intestinal cells by IFA, intestinal tissue was collected from mice, then snap frozen and cryo sections were used for staining the intestinal epithelial cells. Sections were fixed and then blocked. Then, sections were stained with Anti-EpCAM antibody followed by Alexa Fluor® 594 anti-rat IgG staining. Slides were coverslipped with DAPI containing mounting medium and analyzed under fluorescence microscopy. Strong specific signals (positive cells) could be found under fluorescence microscopy.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Mouse intestinal tissue

Primary Incubation

Anti-EpCAM antibody

Blocking Agent

5% FBS in PBS

Secondary Incubation

Alexa Fluor® 594 anti-rat IgG

Tertiary Incubation

NA

Detection

Fluorescence microscopy

Results Summary

Intestinal epithelial cells were stained with very strong and specific signal. The secondary antibody did not cross-react with any other protein of the tissue sample. Thus, we were able to clearly visualize the intestinal epthelia layer.

Additional Notes

Can be used at high dilution rate. Its cost efficient.

Image Gallery

Summary

The Good

Sensitive and can generate strong fluorescent signal

The Bad

None

The Bottom Line

This antibody generated very strong and specific signals and was very good for IFA experiment.

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