Clear Staining Detected with Flow Cytometry

March 25, 2020

Molecular Genetics
University of Toronto
Lab Technician

Overall

Quality of Results

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Company:

Abcam

Product Name:

EpCAM antibody [G8.8] (Allophycocyanin)

Catalog Number:

ab95641

Clone Number:

G8.8

Used for isolating EpCAM+ epithelial cells from embryonic stomach, pancreas, and intestine via fluorescence activated cell sorting for downstream gene expression analysis via qPCR. Goal was to look at changes in epithelial cell gene expression in a control versus mutant mouse model.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Mouse embryonic gut

Primary Incubation

0.125µg for 10^5-8 cells, incubated on ice for 20 minutes

Blocking Agent

2% FBS

Secondary Incubation

None

Tertiary Incubation

None

Detection

638nm laser (Fluorescence activated cell sorting)

Results Summary

Visualization of APC-conjugated EpCAM staining using flow cytometry demonstrated a clear EpCAM+ population as tested in multiple embryonic organs. qPCR validation performed on EpCAM+ versus EpCAM- cell populations demonstrates strong relative enrichment of epithelial markers (i.e. Ecad) and lack of enrichment of non-epithelial cell markers (i.e. Vegf).

DOI or PMID #

N/A

Additional Notes

Image shows live embryonic cells stained with EpCAM-APC, with APC signal strength shown on the vertical axis

Related Categories

Image Gallery

Summary

The Good

Antibody allows for a straight forward staining procedure to produce clean results

The Bad

No problems noted after 2-3 years of use

The Bottom Line

Reliable and convenient APC-conjugated antibody that allows for clean gating/sorting of epithelial cells via fluorescence activated cell sorting.

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