Using CD11b FITC Antibody on Human PBMCS

July 20, 2018

Anaesthesiology
LMU Munich
Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Biolegend

Product Name:

FITC anti-human CD11b Antibody

Catalog Number:

301329

We used this antibody in a panel in order to determine the activation status of certain lymphocyte subpopulations in primary human PBMCs from tumor patients.

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 2 milllion cells)

Blocking Agent

Tru Stain FcX (BioLegend), 10 Mins room temperature (5µl/100µl)

Secondary Incubation

2µl of antibody on 500000 cells, 30 mins on ice in the dark (antibody), wash three times afterwards with 2ml of FACS buffer

Tertiary Incubation

None

Detection

Flow Cytometry

Results Summary

This antibody clearly stains CD11b positive lymphocytes in many subpopulations such as Monocytes etc. In our experiments, the fluorescent intensity was also sufficient to distinguish different expression intensities of this Receptor by MFI.

DOI or PMID #

N/A

Additional Notes

Be sure to always block with a blocking reagent due to unselective binding of the antibodies to monocytes due to Fc Receptors. Due to this matter, always compare this antibody to the corresponding isotype.

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Summary

The Good

Easy to use, concentration mentioned on the tube works great

The Bad

None

The Bottom Line

This antibody reliably stains CD11b. Results are reproducible and coherent. Easy to use, rather inexpensive and easy to use antibody to determine lymphocyte activation.

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