Good Assay Kit To Visualize Phosphatase Tagged Molecules

Johns Hopkins University
Wilmer
Lab Technologist

Overall

Quality of Results

Ease-of-Optimization

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Company:

KPL, a SeraCare company

Product Name:

HistoMark® RED

Catalog Number:

55-69-00

In my studies, I needed to visualize genetically β-galactosidase tagged molecules in mouse tissue with a fluorescent marker in conjunction with other markers. This assay kit allowed me to do that using the Biotin + streptavidin-phosphatase antibody system.

Experimental Design and Results Summary

Application

Immunohistochemistry

Starting Material

Mouse retina tissue

Protocol Overview

Fixed retina tissue is blocked (1% Goat Serum, 0.3% Triton in PBS for 1 hour at room temperature) and then stained with 1:40 biotinylated primary antibody in blocking solution and incubate at 4°C overnight. The tissues were then stained with streptavidin-phosphatase the same was and washed with PBS. The PhThaloRed solution is mixed with the substrate solution and added to the tissue for a 20 minutes stain at room temperature.

Tips

Only prepare Histomark solution right before use.

Results Summary

Though complicated, this stain was the only way we have been able to successfully fluorescently visualize our target cells that are tagged with β-galactosidase.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Works well in mouse tissue

The Bad

None

The Bottom Line

This assay kit works well for fluorescently tagging any phosphatase bound molecules in mouse tissue.

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