Staining Of PBMCs With CD56 FITC

August 02, 2017

Anaesthesiology
LMU Munich
Researcher

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Company:

Biolegend

Product Name:

FITC anti-human CD56 (NCAM) Antibody

Catalog Number:

362545

In order to investigate the CD56 expression in healthy/disease PBMCs, this antibody was included in a panel in order to identify NK and NK T-cells.

Experimental Design and Results Summary

Applications

Flow Cytometry

Sample

Primary human (Ficoll-Isolated) cells

Primary Incubation

10 mins room temperature (Fc Receptor blocking, 5µl for a 100µl cell suspension containing 2 milllion cells)

Blocking Agent

TruStain FcX blocking solution (Biolegend)

Secondary Incubation

2µl of antibody on 500.000 cells, 30 mins on ice in the dark, afterwards wash two times with 2ml of FACS buffer

Tertiary Incubation

None

Detection

Flow Cytometry

Results Summary

This antibody stains CD56 positive cells although the staining results seem to be a little weak. Because of the rather low fluorescence of FITC, for detection of differences in expression a brighter fluorophore should be used. However, no specific staining was observed. Be sure to block with a blocking agent before measuring PBMC preparations via FACS.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

No unspecific background observed

The Bad

Rather low fluorescence, expensive

The Bottom Line

Usable antibody, although detection of expressional changes should be investigated with a brighter fluorophore. Ok for detection of CD56 positive cells. Rather expensive

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