Staining Human PBMCs With HLA-DR PE

August 01, 2017

Anaesthesiology
LMU Munich
Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Biolegend

Product Name:

PE anti-human HLA-DR Antibody

Catalog Number:

361605

This antibody was used in a panel to monitor immunocompetence in intensive care patients together with T-cell, Monocyte and Granulocyte markers.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Primary human PBMCs (Ficoll-Isolated)

Primary Incubation

10 mins room temperature (Fc Receptor blocking, 5µl for a 100µl cell suspension containing 500.000 milllion cells)

Blocking Agent

Tru Stain FcX (BioLegend)

Secondary Incubation

2µl of antibody on 500000 million cells, 30 mins on ice in the dark, wash two times afterwards with 2ml of FACS buffer (PBS with 1%BSA)

Tertiary Incubation

None

Detection

Flow Cytometry

Results Summary

This antibody detects HLA-positive cells without unselective background staining after blocking. The fluorescent brightness of this PE-antibody conjugation is sufficient to distinguish between HLA-DR expression levels on different disease states/patients. Be sure to sufficiently block Fc Receptors when working with human PBMCs.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Easy to use, reliable, not many cells needed, no unspecific staining observed

The Bad

Rather expensive

The Bottom Line

This antibody works quite well in a panel monitoring the immunocompetence of patients. Because of the fluorescent power of PE one is able to detect changes in surface expression levels of this receptor. This antibody produces reliable results.

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