Labeling Endothelial Cells In FFPE Tissue Sections

Univesity of Texas Health Science Center
Translational & Vascular Biology
Assistant Professor

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Company:

Molecular Probes (Thermo Fisher Scientific)

Product Name:

Isolectin GS-IB4 Alexa Fluor 647 conjugate

Catalog Number:

I32450

Marking specific cell types in tissue section has always been a challenge. Isolectin-AF647 conjugate comes handy to mark endothelial cells and vasculatures in tissue sections. In addition to ECs, it can bind to other cell types like microglial cells and neuronal cells.

Experimental Design and Results Summary

Application

Immunofluorescence

Starting Material

Fixed mouse tissue sections

Protocol Overview

Block rehydrated FFPE tissue sections with 3% BSA in PBS for 30 min and then incubate with Isolectin-AF647 (1:200-1:500) for 2 hours at RT. Wash three times with 3% BSA and counter stain with Hoechst 33342 dye. We have included Isolectin-AF647 with primary and secondary antibodies when we stain the section with antibodies. Always works. Did not encounter nonspecific staining.

Tips

Do not thaw and refreeze. It will lead to loss of binding.

Results Summary

Mouse heart section was incubated with 1:200 diluted isolectin-AF647 (crimson color) for 2 hours at RT and then washed 3X with PBS. The section was simultaneously stained with alpha-actinin (green) and Hoechst 33342 dye (blue).

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

No background staining and works all the time

The Bad

In addition to endothelial cells, it binds to neuronal and immune cells.

The Bottom Line

Highly reliable reagent to mark endothelial cells in heart and retina.

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